GENBIO 1.pdf

GENERAL BIOLOGY 1 Sir Ran Abarcar 1ST SEMESTER: 1ST QUARTER rooms where monks lived, so he called them “cells”, which he used MICROSCOPY...

GENERAL BIOLOGY 1 Sir Ran Abarcar 1ST SEMESTER: 1ST QUARTER rooms where monks lived, so he called them “cells”, which he used MICROSCOPY to describe the smallest. - technical field of using microscopes to view objects and areas of objects that can’t be seen with the 5 TYPES OF MICROSCOPE naked eye. 1) Compound Light Microscope MAKING A WET MOUNT collect from - light that passes through one or · container sample in the put the sample more lenses to produce an you slide using a dropper MICROSCOPE · you put coverslip so that , the objective lens won't enlarged image of a specimen. get wet - an instrument that produces an · secure the slide in -consists of ocular lenses the stage w/ stage clips enlarged image of an object. (binocular eyepiece) and objective - Biologists use microscopes to study lenses. cells, cell parts and organisms that S -max 1000x, it magnifies small are too small to be seen with the objects naked eye. yeast Phytoplankton - known as resume species (not good in aquatic) 2) Electron Microscope - has horns Who invented the Microscope? - forms an image of a specimen Anton Van Leeuwenhoek using a beam of electrons rather - Dutch Merchant who learned how than light. to grind lenses to make simple - Resolution: a measure of the clarity microscopes (have only one lens). - of an image; how clear the details - ”Father of Microscopy” and built are over 500 microscopes. - Resolution is the limiting doctor to a - First to discover microorganisms light microscope since the greater under a microscope by observing a the magnification is, the less it is drop of pond water filled with life. able to resolve the image. At He called them “tiny molecules”. magnifications beyond 2,000x, the image becomes blurry, but electron Robert Hooke microscopes can be used at - Improved the microscope greater magnification. - Used a microscope to observe a - black and white thin slice of cork. The spaces he - 100 , 000 x saw reminded him of the small specimen is non-living already preserve - UST-SHS 11 HEALTH ALLIED-4 Batch 2023-2024 GENERAL BIOLOGY 1 Sir Ran Abarcar 1ST SEMESTER: 1ST QUARTER - detects electrons that passed Characteristics of Electron Microscope: through the sample=2D image - A beam of electrons is used to - Images are black and white produce an enlarged image of the - uses an electron beam to study the specimen (does not use light) internal structure of thinly sectioned - This electron beam and the specimens. specimen must be placed inside a - Magnets guide the beam of vacuum chamber so that the electrons toward the specimen, electrons in the beam do not and image is produced to view. bounce off gas molecules in the air. - Magnification is up to 200,000 - Since living things cannot survive in times. a vacuum, the electron microscope -pass through cannot be used to view living cells. 3) Atomic Force Microscope - Much more powerful than light - High resolution type of scanning microscopes probe microscopy 7 to the bending of light waves around an obstacle - 1000x better than optical diffraction a) Scanning Electron Microscope limit (SEM) - Uses a stylus in order to produce an - detects reflected electrons= 3D image image - 3D image - images are black and white - preserve or alive - The specimen is sprayed with a fine 4) Confocal Scanning Laser metal coating (not sliced to view as - Increasing optical resolution & in the TEM) contrast of an micrograph by - As a beam of electrons is passed means of using a spatial pinhole to over the specimen’s surface, the block out-of-focus light in image metal coating emits a shower of formation electrons, and a 3D image of the - Fluorescent dice specimen’s surface is produced to - Commonly used in microbial view. ecology studies - bounce back - specimen is subjectiveto dyes and then views through laser contaminated when there is variety of colors b) Transmission Electron Microscope (TEM) UST-SHS 11 HEALTH ALLIED-4 Batch 2023-2024 GENERAL BIOLOGY 1 Sir Ran Abarcar 1ST SEMESTER: 1ST QUARTER 5) Stereoscope Stage - Where one puts - also known as dissecting the slide to view microscope (dissect small compounds) Stage Clips - Holds slide in Max 100X big distance for the researcher to put. has specimen in thestage lens-stage place PARTS OF MICROSCOPE Revolving - Rotates 360 nosepiece degrees - Change between objectives Ocular Lens - Usually 10x/views the specimen - binocular Arm - Connects base Objective Lenses -¾ to the back of Magnification microscope Scanner: 4x - Supports and Low Power holds the Objectives: 10x microscope High Power Objectives: 40x Iris Diaphragm - Adjusts the Oil Immersion amount of light Objectives:100x that hits the slide from the Coarse - Up and down light source Adjustment Knob (Scanner, LPO, HPO, OIO) Light Source - Where light comes from to Fine Adjustment - Better resolution see image Knob (HPO,OIO) UST-SHS 11 HEALTH ALLIED-4 Batch 2023-2024 GENERAL BIOLOGY 1 Sir Ran Abarcar 1ST SEMESTER: 1ST QUARTER Eyepiece Lens= 5x Base - bottom of the Field Number= 26mm microscope, Low power Objective= 10x used for support. Total Magnification of Lens: Total magnification (x)= (Objective TOTAL MAGNIFICATION OF LENS Magnification)(Eyepiece Magnification) - measure of how much the image is 26𝑚𝑚 26𝑚𝑚 enlarged Solution: (5𝑥)(10𝑥) = 50𝑥 = 0.52 mm - Total magnification (x)= (Objective Magnification)(Eyepiece Example #2: 10x eyepiece lens 22mm low Magnification) power objective Eyepiece Lens= 10x Field Number= 22mm Objective Eyepiece Total Lens Magnific Low power Objective= 10x ation 22𝑚𝑚 22𝑚𝑚 Solution: = = 0.22 mm Scanner 4x 10x 40x (10)(10𝑥) 100𝑥 Low Power 10x 10x 100x Example #3: 30x eyepiece lens FN of 7mm High Power 40x 10x 400x Scanner Eyepiece Lens= 30x Oil 100x 10x 1000x Immersion Field Number= 7mm Scanner= 4x 7𝑚𝑚 22𝑚𝑚 Solution: (30𝑥)(4𝑥) = 100𝑥 = 0.06 mm FIELD OF VIEW - Use millimeter (mm) scale - LPO & Scanner MICROMETRY Equation: -measurement of dimensions of an object 𝐹𝑖𝑒𝑙𝑑 𝑁𝑢𝑚𝑏𝑒𝑟 F.O.V= 𝑇𝑜𝑡𝑎𝑙 𝑀𝑎𝑔𝑛𝑖𝑓𝑖𝑐𝑎𝑡𝑖𝑜𝑛 being observed under microscope - Unit: Microns (µm), also known as Example #1: 5x eyepiece lens FN of 26mm micrometers are a length of measurement Low Power Objective UST-SHS 11 HEALTH ALLIED-4 Batch 2023-2024 GENERAL BIOLOGY 1 Sir Ran Abarcar 1ST SEMESTER: 1ST QUARTER equal to one millionth of a meter. (1,000µm - Has a known value (10 microns per =1 mm; 0.000001 mm is equal to 1µm) space/divisions) Technique 1: Estimated Size (ES) STEP 1: CALCULATE FOR Technique 2: Actual Size (AS) CALIBRATION FACTOR 1) Estimated Specimen Size Formula 1: If not fully seen 𝐹𝑖𝑒𝑙𝑑 𝑜𝑓 𝑉𝑖𝑒𝑤 ES= (𝑠𝑡𝑎𝑔𝑒 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟) (10µ𝑚) 𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑆𝑝𝑒𝑐𝑖𝑚𝑒𝑛 CF= 𝑜𝑐𝑢𝑙𝑎𝑟 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟 𝑢𝑛𝑖𝑡 or CF= (𝑆𝑀) (10µ𝑚) Example #1: 𝑂𝑀 Given– 2mm Example: Number of Specimen–3 (16 𝑠𝑡𝑎𝑔𝑒 𝑢𝑛𝑖𝑡𝑠) (10µ𝑚) 670 um CF= Answer: ⅔ or 0.67µm X1000 = 20 𝑜𝑐𝑢𝑙𝑎𝑟 𝑢𝑛𝑖𝑡𝑠 CF=8 µ𝑚 2) Actual Specimen Size Steps: Formula 2: If fully seen (𝑠𝑡𝑎𝑔𝑒 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟) (𝑆𝑀) a) Calibration Factor CF= 𝑜𝑐𝑢𝑙𝑎𝑟 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟 𝑢𝑛𝑖𝑡 or CF= 𝑂𝑀 b) Actual Specimen Size Example: 1000 µ𝑚 CF= CALIBRATION FACTOR 40 𝑜𝑐𝑢𝑙𝑎𝑟 𝑢𝑛𝑖𝑡𝑠 CF=25 µ𝑚 - Calibrate the ocular micrometer (unknown value) using stage micrometer (known value). STEP 2: ACTUAL SPECIMEN SIZE OCULAR MICROMETER Formula: AS=(Calibration Factor) (Units - Dimensions won’t change even if occupied) the objective lens is switched. - has value Example: an unknown CF= 8 µ𝑚 STAGE MICROMETER AS= (8 µ𝑚)(74 units) - Dimensions change based on the = 592 µ𝑚 power of the objective lens used. UST-SHS 11 HEALTH ALLIED-4 Batch 2023-2024