Bon Secours Mercy Health Richmond-Body Fluid PDF

Bon Secours Mercy Health Richmond HealthPartners Laboratories Title: Body Fluid Counts and Differentials – Automated and Manual Number: HEME-0001 Revision: 8.0 Approval Date: 6/12/2024 Approved By: Clifton Bridges Laboratory Director SMH RGH (CLB...

Bon Secours Mercy Health Richmond HealthPartners Laboratories Title: Body Fluid Counts and Differentials – Automated and Manual Number: HEME-0001 Revision: 8.0 Approval Date: 6/12/2024 Approved By: Clifton Bridges Laboratory Director SMH RGH (CLBRIDGE) Review Date: 6/12/2024 Reviewed By: Clifton Bridges Laboratory Director SMH RGH (CLBRIDGE) HEME-0001 Body Fluid Counts and Differentials – Automated and Manual Copy of version 8.0 (approved and current) Last Approval or Uncontrolled Copy printed on 12/12/2024 11:08 6/12/2024 Periodic Review Completed AM Next Periodic Review Leetta Bateman Lead MLS 6/12/2025 Printed By Needed On or Before (lcbatema) Effective Date 6/12/2024 Organization Richmond Market Approval and Periodic Review Signatures y p M co11:08 Type Description Date Version Performed By Notes Approval Lab Director 6/12/2024 8.0 Clifton Bridges Laboratory Director SMH RGH (CLBRIDGE) A Laboratory Approval Director SMC SVMC 6/12/2024 8.0 d Mark Williams Laboratory Director (WILL1050) e 24 Laboratory l l 0 o Approval Director 5/7/2024 8.0 Jack Wilson Laboratory Director SPEC RCH (JJWILSON) 2 2/ r 12/1 SPEC RCH t n f Approval Laboratory Director 5/6/2024 8.0 o c ta s o SECC n rren U Cu Angie DUONG Laboratory Director (ADUONG) Laboratory Approval Director 5/6/2024 8.0 Jeffery Green MD (JTGREEN) SFMC WEC Laboratory Approval Director 5/6/2024 8.0 Lindsey Matthews Laboratory Director (MATT0083) CEC Laboratory Approval Director 5/6/2024 8.0 MRMC CoraGabriela UramTuculescu Laboratory Director (URAM0000) Owner Approval 5/6/2024 8.0 Approval Thuy Sandy Pham Education Coordinator (101706) New Lab Approval Director 3/6/2024 7.0 Approval  CoraGabriela UramTuculescu Laboratory Director (URAM0000) New Lab Approval Director 2/24/2024 7.0 Mark Williams Laboratory Director (WILL1050) Approval New Lab Approval Director 11/7/2023 7.0 Lindsey Matthews Laboratory Director (MATT0083) Approval Periodic Designated 10/30/2023 7.0 review Reviewer Thuy Sandy Pham Education Coordinator (101706) Recorded on 5/9/2023 by Thuy Sandy Minakowski Education Periodic Coordinator (101706) review Designated when document Captured 12/12/2022 7.0 Thuy Sandy Pham Education Coordinator Reviewer added to Document outside Control (previous MediaLab system of record: Master Control y p M Document Records) Recorded on 5/9/2023 Approval Captured Lab Director 12/9/2022 7.0 o c :0 Clifton Bridges Laboratory Director (CLBRIDGE) 8 A by Thuy Sandy Minakowski Education Coordinator (101706) when document outside MediaLab d e 24 11 added to Document Control (previous l l 0 system of record: Master Control Document Records) t r o 2 / 12 / 2 Approvals and periodic reviews that occurred before this document was added to Document Control may not be listed. Version History o n 1 of Date Added Date Effective Date Retired c Version Status Type 8.0 Approved and Current Major revision as 5/1/2024 6/12/2024 Indefinite 7.0 Retired n First version in Documentn e t Control 5/9/2023 12/12/2022 6/12/2024 U C ur r  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual PRINCIPLE Routine body fluid analysis consists of gross examination for color and clarity, a total RBC count, a total nucleated cell count (including WBC), a differential count, and a search for abnormal cells and crystals. Validated fluid sample types are analyzed on the Sysmex XN Hematology analyzers to obtain a total WBC and RBC count. Specimens must be run in duplicate and produce similar results. If similar results are not produced, manual counts must be performed. Manual and lower cell counts are performed using a Neubauer hemacytometer and isotonic saline (Cellpack), as needed. Manual differential counts are performed on a Wright's stained cytocentrifuged slide. SPECIMEN y p M Specimen Type Order Code o sequentially o Specimen Collection c :0 A CSF is submitted in 3 sterile tube at the time of collection and labeled 8 Cerebral Spinal Fluid (CSF) SFCCT d 11 ▪ Tube 1 – Chemical and Immunologic studies ▪ Tube 2 – Microbiologic examination e 24 o l ▪ Tube 3 – Cell Count and Cytological examination l 0 CSF will also be acceptable in any non-additive container Serosal Fluid FCCT o t r 12/1o peritoneal and pericardial. 2/ 2 Fluids from the main body cavities obtained through centesis: pleural, o n f Collect in an EDTA tube or non-additive container, with an ideal volume of 3-7 mL o o o o c ta Collect in an EDTA tube or non-additive container, with an ideal s volume of 3-7 mL n rren Crystal analysis Synovial (Joint) Fluid SNCCT U Cu ▪ Collect in a Red top tube or non-additive container ▪ Avoid the use of EDTA as it may cause interference with crystal analysis ▪ If EDTA is the only sample available, perform the crystal analysis and attached the following to a positive result, ‘Analysis performed on EDTA, evaluate results with caution.’ o To include: Bronchial Washings, Amniotic Fluid, Jackson Pratt Body Fluids from all Drainage, Peritoneal Dialysis Fluids other sites or o These fluid types are NOT Approved for performance on the Sysmex interventions analyzers and must be performed manually. o Due to the rapid degradation of these specimens, it is appropriate to perform a cell count using the hemocytometer immediately. Hematomas and o If the performing tech feels the specimen is ordered incorrectly, it is other unusual fluids appropriate to elevate to the pathologist/pathologist on-call o It is appropriate to hold the results of the cell count while a pathologist reviews the case and consults with the provider. Page 1 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 1 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual Handling Conditions When a specimen is submitted in only one container/tube and multiple tests are requested: The technical staff is to refer to the Microbiology Aseptic Technique Procedure to aliquot sample into appropriate containers. The originating facility is to maintain proper supplies and utilize technical personnel to ensure the integrity of the additional testing in the SMH core automated, microbiology, cytology, or reference laboratories. Sterility is required in this process. Technical staff must perform this pour-off or aliquot. Use a freshly wiped down area for the transfer. 1. Label sterile aliquot container with the proper patient label. 2. Write your tech code on label. y 3. Using a sterile transfer pipette and the Microbiology Aseptic Technique Procedure, aliquot fluid to the labeled container(s), taking care not to compromise the sterility of the fluid. Volume will vary based on testing needs and amount submitted. p M o 4. Track appropriately to the SMH Core Microbiology, Cytology, or Reference Lab. c :0 8 RCH does not perform any fluid testing and all samples are immediately tracked to SMH. A d 11 If volume submitted is determined to be inadequate for all testing orders, the testing site e 24 performed first. l l technical staff is to communicate with the caregiver to prioritize and triage testing to be 0 Specimen Storage and Stability t r 12/1o 2/ 2 Prompt examination of all fluids is a priority. o CSF samples are handled as a STAT. o on f c ta o Ideally performance is examination within 2 hours of arrival in the lab. n rren s o Attach the following comment to fluids that are performed >8 hours after collection: ▪ SPEDL: translates to “A delay in specimen examination occurred. Please U Cu evaluate results carefully.” Fluid specimens are stored for 7 days at 2-8°C. Fluid cytosmear slides are stored for at least 1 week at room temperature. Unacceptable Specimens All specimens are accepted with no exceptions. If the specimen is improper, ensure that the discrepancy is noted in the patient report. Check samples for clots. If the fluid specimen is partially clotted or has cell clumps or interfering debris on the chamber, attach the code: CLOTF to the cell count results. o CLOTF: translates to “fluid specimen clotted, count may be inaccurate.” QNS Specimens: If volume submitted is judged to be inadequate for all testing orders; the testing site technical staff is to communicate with the caregiver to prioritize and triage tests to be performed first. Testing that cannot be performed may be credited as QNS. Page 2 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 2 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual EQUIPMENT, MATERIALS, AND REAGENTS Equipment and Materials Sysmex XN Hematology Analyzer (see CBC-XN procedure for full details) Neubauer Hemacytometer or Disposable Hemacytometer Microscope Cyto-Tek Centrifuge Cyto-Tek Specimen Chamber Assembly Manual Cell Counter Microscopic Slides Automatic Pipettes, Sterile Disposable Pipettes Wooden Applicator Sticks y p M Petri Dish with Filter Paper Microhematocrit/hematocrit capillary tubes Clean plastic or Sterile Vacutainer with no additive o c :0 8 A Reagent Storage and Stability d e 24 11 Reagent Upopened Stability l l Opened Stability 0 Notes Cellpack – Diluting Fluid for Manual Dilutions Until expiration at 15-30°C tr o 2/ 1 2 30 days at 15-30°C /2 Background checks on the Diluting Fluid are performed daily in the CBC mode, document Wright-Giemsa Stain Pack Until expiration n o so f 1 60 days at 15-30°C results in QCE New Methylene Blue n c Until expiration at n t a This stain may assist in the differentiation between RBCs and (Loeffler’s) UC 15-30°C r re 60 days at 15-30°C WCBs on the hemocytometer. If utilized, record use with the u report. See Procedure Notes for Directions for Use Ensure to keep container tightly Hyaluronidase 5 years at -20°C per closed when not in use. See manufacturer Procedure Notes for Directions for Use QUALITY CONTROL & BACKGROUND COUNTS ON DILUTING FLUID QC Material QC Frequency Unopened Stability Opened Stability Sysmex XN Check BF 2 levels once every 24 84 days at 2-8°C 30 days at 2-8°C Automated QC hours One level for each 8 hours Quantimetrix Spinalscopics 6 months at 2-8°C of patient testing Until expiration at 2-8°C Manual QC 1 month at 18-25°C (refer to QCE for schedule) Cellpack Background check Until expiration at 15-30°C 30 days at 15-30°C Diluting Fluid performed daily Page 3 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 3 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual QC Material Sysmex XN Check BF – Automated Body Fluid Controls o Levels 1 and 2 o Do not store frozen or freeze the control o Storage outside the recommended temperature range causes damage to the product. Deterioration is suspected when the mean of a control result is not within the assay expected ranges after appropriate troubleshooting. Do no use vials; discard any vials that are confirmed to be damaged by the supervisor. o If deterioration is suspected, call the Sysmex Technical Assistance Center and contact department supervisor/manager. Quantimetrix Spinalscopics – Manual Fluid Cell Count Controls o Levels 1 and 2 o Discard the control vial if there is any evidence of microbial contamination. y p M Cellpack – Diluting Fluid used for Daily Background Check and Manual Dilutions All Controls must: o c :0 8 A Be reviewed and found acceptable before resuming patient testing d e 24 11 Be entered into Unity; all responses, including any failures l l Have documentation of corrective action taken in Unity when found unacceptable 0 Background Checks must: t r 12/1o 2/ 2 Be reviewed and found acceptable before using Be entered into QCE o on f Performing QC – Sysmex XN Check BF (Automated) c ta Have documentation of corrective action taken when found unacceptable n rren s U Cu 1. Remove a vial of XN Check BF control from the refrigerator and equilibrate to room temperature (15-30°C) for 15 minutes before use. 2. Analyze the control in the instrument QC Analysis Mode according to the Sysmex XN series Instructions for Use. The pierceable septum in the vial cap allows for closed tube analysis. 3. Switch to Manual Mode; on the control menu, click the change analysis mode button and select Body Fluid. Click OK. 4. Wait for the BF BACKGROUNDCHECK to complete. Print out background count and attach/file with the manual fluid worksheet. To print: Highlight the background count, then Click Validate button. Press Output button. Then select GP printer. 5. Mix the vial by end-to-end inversion until all red blood cells are completely resuspended (approximately 20 inversions). Do not place on a mechanical mixer. 6. Place XN CHECK BF Level 1 onto the sample tube holder. 7. Click the manual analysis button on the control menu and ensure Read ID is checked and Cap Open is unchecked. Click OK. 8. Press the start switch on the analyzer. After aspiration, the tube holder will slide out. Remove the tube. 9. To process the XN CHECK BF Level 2, wait until the status light is solid green and then continue. Page 4 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 4 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual 10. Return controls to the refrigerator immediately after use. 11. Review controls in the instrument QC files to determine acceptability. Controls must be within acceptable tolerance limits for patient reporting. 12. If controls are unacceptable, troubleshooting and corrective actions must be performed. 13. Document control acceptability or unacceptability, troubleshooting and any corrective actions in Unity. Performing QC – Quantimetrix Spinalscopics (Manual) 1. Remove the control from the fridge and allow to come to room temperature (18-25°C) for at least 15 minutes, depending on the volume remaining in the control vial. a. If removing a new vial, replace the cap with the bulb dropper cap times and by gently squeezing and releasing the dropper at least 10 times. Avoid foaming. y 2. Immediately prior to use, mix the controls thoroughly by gently inverting the bottles several p M o 3. Using the dropper, charge both sides of the hemacytometer chamber. Immediately recap the c :0 8 A control. Allow the cells to settle by placing the hemacytometer in a humidified chamber for 10 minutes before counting. d 11 4. Count the RBCs and WBCs in the 9 large squares of the hemacytometer. For each side, calculate e 24 the total number of cells as follows: Number of cells counted = Total cells/µL l l 0 0.9 t r 12/1o 5. The total cells counted on each side of the chamber must agree within 20%.2/ 2 6. Document control counts and calculations on the fluid worksheet or QC Checkoff sheet. A blank worksheet may be printed from LIS. o on f 7. The average of the total cells counted on each side is entered in Unity. c ta n rren s Performing a Background Check – using Cellpack (Diluting Fluid) Cellpack and the date. U Cu 1. Aliquot a small amount of Cellpack into a non-additive tube. Properly label the tube with 2. Run the diluting fluid in the CBC mode. The aspiration sensor may need to be turned off to allow for sampling. 3. Acceptable Range o WBC: ≤0.10 x103/µL o RBC: ≤0.02 x106/µL 4. Print a copy of the background count and document in QCE. 5. The Cellpack aliquot is good for 24 hours and may be used for body fluid dilutions. A new aliquot must be made daily for the background counts. Page 5 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 5 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual y p M o c :0 8 A d e 24 11 l l 0 t r 12/1o 2/ 2 o on f c ta n rren s U Cu Page 6 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 6 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual TESTING PROCEDURE Part 1: Gross Examination a. Note the Color and Appearance of the fluid. Color: Colorless, Yellow, Red, Straw, Amber, etc. o A color may be free text if needed. o Do not use Xanthochromic as a color. Appearance: Clear, Hazy, Cloudy, Bloody, Turbid, etc. b. CSF Color: If CSF color is anything other than Colorless, centrifuge sample and examine supernatant. LIS allows for documentation, in defined fields, of unspun and spun color: Unspun Color: ______ Spun Color:_______ c. Check for Clots. Do not run a clotted fluid on the instrument. Note: If fluid is clotted, attach CLOTF to WBC results. y p M Part 2: Fluid Counting – Automated and Manual Notes o c :0 8 A Automated Counts: Are only approved for CSF (other than clear and colorless), serosal, and synovial fluid. d e 24 11 Manual Counts: All other fluids are to be performed manually (e.g. bronchial washings, l l bronchial lavage, peritoneal dialysis, amniotic and Jackson Pratt drainage fluids). If fluid volume is limited, then a manual count should be performed. 0 t r 12/1o 2/ 2 Large numbers of RBCs can make manual WBC counting difficult. Dilution of the sample with Cellpack may be performed. Ensure dilution factor is entered into the calculation of cells for obtaining final results. o n f All synovial and viscous fluids must be prepared with the addition of hyaluronidase prior to o c ta s counting. See Directions for Use in Procedure Notes. n rren If using New Methylene Blue Stain (Loeffler’s) see Directions for Use in Procedure Notes. Automated Fluid Counts – XN: U Cu 1. Switch instrument to Manual Mode, Click Change Analysis button, Select Body Fluid, Select OK. 2. The XN will perform a Background Check automatically once the Body Fluid analysis mode is selected. a. A Background Check (autorinse) is performed before every fluid sampling to rule out any carryover from prior whole blood samples. Review for acceptability and attached to the manual worksheet for the fluid count and retain for two years per HPL policy. b. Print out the background count and attach/file with the manual fluid worksheet. Must not be in last 20 Mode. To print: Highlight the background count > Click Validate > Press Output > select GP printer. 3. Review the Manual Analysis Screen a. Enter Sample No., if the instrument is unable to read the barcode. b. If the cap is removed, ensure the Cap Open is selected (a check mark in the box). c. Once review ID completed, select OK to proceed. 4. Thoroughly mix the fluid immediately prior to sampling to ensure sample is well mixed. a. Note: Excess or aggressive mixing can disrupt blood cells, invalidating results Page 7 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 7 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual b. If needed, transfer fluid to a properly labeled tube for sampling. 5. Place sample on the instrument and Press the blue Start button to begin analysis. a. All fluids must be performed in duplicate. 6. Review the TNC-BF# and RBC-BF. a. If result is within the Linearity (see below), repeat sample. b. If result is outside the Linearity, dilute fluid with Cellpack and run in duplicate. i. Note: Ensure to include dilution factor in final calculation. 7. The duplicate run for WBC and RBC must agree within the stated criteria a. 0.05-0.15 must agree within 20% b. >0.15 must agree within 15% c. If count does not agree, perform testing again 8. If any of the following occur, a manual count must be performed: a. TC-BF is 0.003 – endpoint x 106/L 0.003 – 5,000 x 106/L e Reporting Automated Counts from Instrument C uUrr Report results as whole numbers and record on worksheet: TNC-BF/WBC: move decimal 3 places to the right. o 0.060 and 0.050 would be reported as 55 o 1.200 and 1.400 would be reported as 1300 RBC: move decimal 6 places to the right. o 0.03 and 0.04 would be reported as 35,000 o 0.18 and 0.20 would be reported as 190,000 Note: RBC counts >100 are to be reported as >100 on all fluids, with the exception of CSF. Manual Fluid Cell Counts 1. Thoroughly mix the fluid and load, being careful to not overfill, both side of the Neubauer Hemacytometer with a capillary tube. a. See Procedure Notes for Directions for Use. b. Hemacytometer should be clean, free of scratches, and all lines clearly visible. c. Optional: Rinse capillary tube with Methylene Blue, see Procedure Notes for details. 2. Allow fluid to settle in a Petri dish continuing wet filter paper, for 10 minutes. Page 8 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 8 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual a. Settling allows the cells to stop drifting and to be in the same plane of focus. b. Do not allow fluid to settle too long, this may cause the sample to dry out, concentrating the cells. 3. If the fluid has no overlapping cells, count the undiluted fluid. Counts between both sides of the Hemacytometer must agree within 20%. Record results on fluid worksheet. a. Cell counts must be performed in duplicate (count both sides of the Hemacytometer) 4. If cells are too numerous to count, dilute with Cellpack. Reload the Hemacytometer and proceed with count. a. Suggested Dilutions i. 1:10 dilution for slightly cloudy fluids ii. 1:20 dilution for moderately cloudy fluids iii. 1:100 dilution for very cloudy and grossly bloody fluids 5. b. Note: Ensure to include dilution factor in final calculation y p M When counting any of the 9 large squares (indicated by orange box below) on each side of the Hemacytometer, use the following formula: 6. a. Total cell/µL3 = # of cells counted x dilution factor # of large squared counted x 0.1 o c :0 8 A When counting any of the 25 small squares (indicated by green box below) on each side of the Hemacytometer, use the following formula: a. Total cell/µL3 = # of cells counts x dilution factor d e 24 11 # of small squared counted x 0.004 l l 0 t r 12/1o 2/ 2 o on f c ta n rren s U Cu Page 9 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 9 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual Part 3: Smear Preparation 1. Prepare a cytosmear slide for the following: a. Any white cells/nucleated cells are present in a fluid, other than CSF. b. ≥5 white cells/nucleated cells are present in CSF. 2. Cytocentrifuge the fluid at approximately 1500 RCF for about 5 minutes. 3. Stain slide with Wright’s stain. 4. Guidelines to preparing a slide: a. Label slide with patient name, accession number, date, and tech code b. Assemble to the sample chamber, filter card, and slide. c. Depending on the appearance of the fluid, a varied amount of sample may be used. i. Clear fluid: 5-10 drops ii. Cloudy fluid: 1-3 drops iii. Bloody fluid: 5-10 drops of a 1:5 saline dilution y p M o d. Adding 1-2 drops of 22% bovine serum albumin to the bottom of the sample chamber c :0 8 A may help reduce the cell distortion and increase yield (with the exception of synovial fluid). e. RCFs may be reduced if WBC appear to be fragile. d e 24 11 f. Once chamber is filled, cap and place in cytopsin. l l 0 Part 4: Fluid Differential Fluid Type t r 12/1o 2/ 2 Perform a 100 cell differential, when possible, using the proper fluid keyboard. GUI Keyboard CSF Fluid o on f SFG FG c ta Synovial Fluid n rren If a 100 cell differential is not possible: s SYNFG U Cu o Set count to a lesser number under ‘Set Count’, or o Use the ‘Term’ key on the diff keyboard to terminate the count Part 5: Pathologist Review (if indicated) All slides showing abnormal, unusual or suspicious results must be reviewed by a pathologist. Refer to HPL Pathologist Review procedure and completed a Path Review form for submission. Stats require delivery and/or call to the pathologist for immediate review, if indicated by nurse or physician. REPORTING RESULTS Reference Ranges: See Hematology Global Test Parameters. Reporting in LIS: Fluid Count o Function: MEM o Worksheet: Page 10 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 10 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual Site Worksheet MRFCC - Fluids and Synovial Fluids MRMC CSF - Spinal Fluids RGH RMANH – All Fluids FFCC - Fluids and Synovial Fluids SFMC SFCSF - Spinal Fluids SFWC FWCSF SMH F – Fluids and Synovial Fluids SF – Spinal Fluids y p M SPEC SPMHEM – All Fluids o c :0 8 A o Fluid Type: LIS Code Fluid Type d e 24 11 PERI Pericardial l l 0 PERT PLEU BROWN Peritoneal Pleural t r 12/1 Bronchial Washing o 2/ 2 JP AMN o on f Jackson Pratt Amniotic CSF PFLD c ta s Cerebral Spinal n rren Paracentesis PTDFL SYN U Cu Peritoneal Dialysis Synovial Free Text if code not available o Fluid Color: LIS Code Color COL Colorless RED Red YEL Yellow STRAW Straw ORNG Orange GRN Green BROWN Brown PINK Pink AMBER Amber Free Text if code not available Page 11 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 11 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual o Fluid Appearance: LIS Code Appearance CLEAR Clear HAZY Hazy CLDY Cloudy TURB Turbid BLDY Bloody Free Text if code not available o Fluid WBC/Total Nucleated Count: Enter Count o Fluid RBC Count: Enter Count Fluid Differential o Select Keyboard in GUI: y p M ▪ FG – Body Fluids ▪ SYNFG – Synovial Fluids ▪ SFG – Spinal Fluids o c :0 8 A o Enter Accession Number. d e 24 11 o Perform Differential: Key on Fluid l SYN l 0CSF Keyboard M Keyboard snseg t r 12/1o Keyboard fsegs 2 / 2Keyboard Sfsegs. , snlym snmon o on f flym fmm Sflym Sfmon J I sneos snoth c ta n rren s feos fothc Sfeos Sfoth o / ‘ U Cu fslin Perform QA Review and Save. flmeso Sfml sfmac Reporting Notes: o Bronchial Washing may show numerous alveolar macrophages. These cells have an eccentric/round nucleus, light blue cytoplasm, a variable number of azurphilic granules in the cytoplasm and appear similar to macrophages seen in pleural or peritoneal fluids. Blue-Black cytoplasmic particles can be seen in cells of smokers. o Bronchial Lining and Mature Squamous cells are contaminants of bronchial lavage samples and indicate proximal sampling. Bronchial cells are cylindrical in shape with eccentric nuclei and a tuft of cilia. Mature Squamous cells are large, flat and contain small nuclei that are not more than 10-20% of the total cytoplasmic volume. o Bronchial Washing specimens do not contain Mesothelial cells. Page 12 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 12 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual PROCEDURAL NOTES Most Common Expected Observations: 1. Abnormal CSF will be found in the following conditions: i. Meningitis vi. Cerebral infarct ii. CNS Hemorrhage vii. Cerebral abscess iii. Multiple Sclerosis vii. Previous lumbar puncture iv. CNS Malignant Conditions ix. Myelography, pneumonencephalography v. Guillain-Barre syndrome x. Post chemotherapy and radiation changes 2. Causes of Pericardial Effusions i. Infections ii. Cardiovascular disease iv. Neoplastic disease v. Trauma y p M iii. Renal Failure and uremia 3. Causes of Pleural Effusions o c :0 8 A i. Infectious Diseases ii. Neoplastic Diseases d 11 v. Gastrointestinal disease e 24 vi. Postmyocardial infarction iii. Trauma iv. Collagen vascular disease l l vii. Pulmonary Embolism, Infarction viii. Chylous effusion 0 4. Causes of Peritoneal Effusions t r 12/1o 2/ 2 i. Congestive Heart Failure ii. Cirrhosis o n f vi. Trauma vii. Pancreatitis o iii. Hypoproteinemia iv. Infections v. Malignancy c ta n rren sviii. Carcinoma ix. Tuberculosis U Cu 5. Causes of Abnormalities in Synovial Fluid i. Acute Inflammation iv. Gout ii. Chronic Inflammation v. Lupus Erythematosus iii. Rheumatoid Arthritis Slide boxes containing cytocentrifuged fluid smears with unusual or abnormal cells and Body Fluid books with colored photos are available in the laboratory at all times for technologist review. CAP photomicrographs are used for annual competency for all performing technologists. Fluid Crystal Analysis: Oxalate or powered EDTA should not be used, as they may produce crystalline structures that resemble monosodium urate. If EDTA is the only sample available for analysis attached the following a positive result ‘Analysis performed on EDTA, evaluated results with caution.’ New Methylene Directions for Use: 1. Aliquot a small sample of body fluid into a sample cup. 2. Place three to four drops of methylene blue stain into a secondary sample cup. Page 13 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 13 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual 3. Take a hematocrit tube and partially fill it with the methylene blue stain. 4. Place edge of hematocrit tube on a piece of gauze to draw the excess stain out of the tube. The hematocrit tube should now be stained blue with no excess liquid. 5. Using the stained hematocrit tube place into the first sample cup containing the aliquot of specimen. Draw enough fluid to fill the hematocrit tube ¾ full to full. Let stand horizontal for approximately 10 minutes to let the cells stain. 6. After 10 minutes of staining is completed, take the stained sample and load onto a clean hemacytometer. 7. Let stand 10 minutes in a humidified chamber prior to reading. Hyaluronidase Directions for Use: 1. Mix viscous fluid well by inversion. y p M 2. Transfer a 1-2mL aliquot of the viscous fluid to a clean plastic or sterile Vacutainer tube. and mix well. 4. Cap aliquot tube and mix well by inversion before analysis. o 3. Using two wooden applicator sticks, transfer a few grains of hyaluronidase to the fluid aliquot c :0 8 A 5. Wait 5 minutes or until aliquot is less viscous. d e 24 11 Hemacytometer Directions for Use: l l 0 Metallized Hemacytometer Hausser Bright-Line Directions for Use t r 12/1o 2/ 2 Bright-Line / Dark-Line Counting Chambers Catalog Numbers: 3100, 3110, 3200, 3500, 1490, 1492, 1475, & 1483 Usage: Cell Counts Cell Depth: 0.100mm +/- 2% (1/10mm) o on f Volume: 0.1 Microliter c ta n rren Ruling Pattern: Improved Neubauer, 1/400 Square mm s U Cu Rulings cover 9 square millimeters. Boundary lines of the Neubauer ruling are the center lines of the groups of three. (These are indicated in the illustration below.) The central square millimeter is ruled into 25 groups of 16 small squares, each group separated by triple lines, the middle one of which is the boundary. The ruled surface is 0.10mm below the cover glass, so that the volume over each of the 16 small squares is.00025 cubic mm. Neubauer Ruling Page 14 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 14 of 15  Bon Secours Mercy Health Richmond HealthPartners Laboratories Body Fluid Counts and Differentials – Automated and Manual HEME-0001 (Version 8.0) Approved and current. Effective starting 6/12/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Hematology Procedure: Body Fluid Counts and Differentials, Automated and Manual The number of cells per cubic millimeter = Number of cells counted per square millimeter X dilution (if used) X 10 The number of cells per milliliter = Number of cells counted per square millimeter X dilution (if used) X 10,000 One (1) Milliliter = 1000 cubic millimeters (cu mm) One (1) Microliter (ul) = One (1) cubic millimeter (cu mm) To clean the counting chamber: After completing the count, remove the cover glass and clean the counting chamber with water or a mild cleaning solution (10% solution of bleach). Dry the counting chamber with a soft cloth or wipe, or rinse with acetone. PROFICIENCY TESTING AND INTERLABS y p M o c :0 A CAP surveys and instrument interlabs are performed at regular frequency to meet regulatory guidelines. 8 After results are submitted, CAP survey slides are retained for review by all technologists who perform d 11 fluid differentials to ensure competency and to ensure that all technologists report microscopic morphology in the same manner. e 24 l l 0 REFERENCES t r 12/1o 2/ 2 1. Kjeldsberg, Carl R. and Knight, Joseph A; Body Fluids. Laboratory Examination of Amniotic, Pathologists Press Chicago IL, 1993. o on f Cerebrospinal, Seminal, Serous & Synovial Fluids 3rd Edition, American Society of Clinical W. B. Saunders Co., Philadelphia 2007. c ta 2. Henry, John Bernard: Clinical Diagnosis and Management by Laboratory Methods. 21st Edition n rren s 3. Miles Inc. Elkhart IN, CYTO-TEK Centrifuge Operating Manual.99A43259 Rev. 10/90 U Cu 4. Ringsrud, K.M., Urinalysis and Body Fluids: A Color Text and Atlas; Mosby Co., St. Louis 1995. 5. American Thoracic Society “Clinical Role of Bronchoalveolar Lavage in Adults with Pulmonary Disease”. 1990. Am REV Respir Dis 1990 142:481-486. 6. Galgan, Blomberg, Cornbleet & Glassy, “Color Atlas of Body Fluids”. College of American Pathologists. 2006. 7. Quantimetrix Spinalscopics package insert. Quantimetrix, CA. 2005. 8. Shidham, V.B, Atkinson, B.F Cytopath Diagnosis of Serous Fluids, Saunders 2007, Elsevier Inc. 9. Editors Galagan, MD, K.G., Blomberg, MD, D., Cornbleet, MD, P.J.C. 10. Glassy, MD, E.F Color Atlas of Body Fluids ; An Illustrated Field Guide Based on Proficiency Testing CAP Hematology and Clinical Microscopy Resource Committee 2006. 11. Husson, Jerry W., MD, and Kjeldsberg, Carl R, MD, Kjeldsberg’s Body Fluid Analysis, American Society for Clinical Pathology, 2015. 12. Sysmex Corporation, Automated Hematology Analyzer XN Instructions for Use and Operator’s Manuals (Current as of January 2018). 13. Sysmex Corporation, Automated Hematology Analyzer XT Instructions for Use and Operator’s Manual. 14. CLSI Analysis of Body Fluids in Clinical Chemistry, C49-A Vol. 26 No. 21. Page 15 of 15 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:08 AM (EST). Page 15 of 15