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Equipment
Lab
| Question | Answer |
|---|---|
| Simple vs compound microscope? | One lens vs two (light scope) |
| Scanning lens magnification? | x2.5 to x4 |
| Intermediate lens magnification? | x10 to x20 |
| High-powered dry lens magnification? | x40 to x45 |
| Oil immersion lens magnification? | x90 to x100 |
| 4 common light microscope objectives? | Scanning, intermediate, high-powered dry, oil immersion |
| What is resolving power? | Closest two objects can be & still appear separate |
| What is chromatic aberration? | Distortion of color in image due to lenses being incorrectly angled |
| Total magnification = | Objective magnification X oculars magnification |
| Polarizing microscope used to? | ID crystals (talc, silica, urates); amyloid stained w/ Congo red |
| What is used to view substances exhibiting double refraction, anisotropism, & birefringence? | Polarizing microscope |
| What is birefringence? | Property of refractive index being dependent on polarization & propagation of light |
| Convert light scope to polarizing by? | Analyzer between specimen & eye, polarizer between light source & specimen |
| Anisotropic or birefringent objects appear? | Bright against dark background |
| Phase-contrast microscope used to? | View unstained, esp. living, specimens, transparent objects |
| Dark-field microscope used to? | Study unstained microorganisms |
| Dark-field microscope works by? | Excluding direct light, only scattered/oblique light, small objects appear larger & luminous |
| Fluorescence microscope works by? | Emitting light λ that's absorbed by substance & reemitted as longer λ |
| Fluorescence scope light source? | Mercury or halogen lamp, exciter filter between light & specimen, barrier filter in eyepiece to absorb UV |
| Primary/autofluorescence means? | Substance naturally fluoresces, e.g. collagen |
| What is a fluorochrome? | Absorb one λ of light & emit another, usu. UV light, include: rhodamine, fluorescein isothiocyanate (FITC, green), & Texas Red, fade quickly |
| What is auramine-rhodamine? | Fluorescent dye for acid-fast bacilli |
| What is thioflavin T? | Fluorescent dye for amyloid |
| Two types of electron microscopy? | Transmission & scanning |
| Transmission microscopy works by? | Specimen transmitting/deflecting electrons; 2D black & white image on fluorescent screen |
| Scanning microscopy works by? | electron beam causes specimen to release electrons, 3D image, great depth of focus |
| Transmission microscopy great for? | Diagnosis of kidney dieases, tumor ID |
| 4 types of microtomes? | Rotary, sliding, ultra, clinical freezing |
| Ultramicrotome section thicknesses? | 0.5 μm plastic for light, 90 nm for EM |
| Rotary microtome mechanism & block type | Block moves past knife, frozens, paraffin, GMA |
| Sliding microtome mechanism & block type | Knife moves past block, large paraffin blocks, celloidin |
| Clinical freezing microtome | Mostly replaced by cryostat, bad for friable tissue, easier free--floating sections, use CO2 to freeze blocks, infectious agent inhalation hazard |
| Wedge-shaped knife used to section? | Paraffin, Carbowax, frozens |
| Planoconcave knife used to section? | Celloidin, blade must be kept wet |
| Average paraffin knife clearance angle? | 3° to 8° |
| What is a cryostat? | Refrigerated chamber containing (usually rotary) microtome |
| Optimum cryostat knife tilt? | 30° |
| Average cryostat operating temp? | -20°C, colder for fat, warmer for brain, liver, spleen, lymph node, endometrial scrapings |
| Slides must be completely dry before deparaffinization because? | Water contaminates xylene, incomplete deparaffinization, white spots in tissue |
| Incubator temperatures? | 37°C, body temp; for enzyme rxns, special stains |
| Fridge/freezer temperatures? | 4°C to 10°C / -20°C |
| Linear stainers | Move slides from one container to next, same time each container |
| Revolving stainers | Move slides from one solution to next, can set times |
| Robotic stainers | Allow total programming, slides can go in containers in any order |
| Diamond knives | Used for v. hard resin |
| Glass knives | Used for resin, e.g. Spurr's, make just before use |
| Disposable knives | Used for paraffin, GMA, frozens, high profile - used w/ firm, difficult tissues, or range of tissues, low profile - Cryostat & soft tissues |
| Steel knives | No longer used in histology, sharpened by hand |
| Bevel angle | aka facet angle, angle on tip of microtome blade |
| Common section thicknesses | Routine 3-5 µm (~1 cell thick), some biopsies 2 µm, brain & nervous 8 µm |
| Cryostat should be cleaned with? | 70% ETOH |
| Cryostat blades should be changed: | Between cases to avoid risk of exposure & cross-contamination |
| How often should pipettes be calibrated? | At least annually |
| A binocular microscope __ | Has two viewing oculars |
| Parfocal lens | Stays in focus when magnification is changed |
| Achromatic lens | Bring two wavelengths (usu. red & blue) into focus in same plane to reduce distortion due to refractive index differences |
| Apochromatic lens | Bring red, blue & green wavelengths into focus in same plane to reduce distortion due to refractive index differences, more expensive , not used in routine histology |
| Why should metals not be used in the microwave oven? | They don't transmit or absorb microwaves, they reflect microwaves, cause sparking. |
| Is microwave radiation ionizing or nonionizing? | Nonionizing |
| Molecules that generate heat when exposed to microwaves are: | Microwave absorbent, not microwave transparent |
| What 3 things should be carefully controlled when using a microwave oven? | Exposure time, solution volume, oven wattage, to avoid heat damage to specimen |
Created by:
CCF
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