fluorescence microscopy

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barrier filters should not ...

absorb the fluorescent wavelength

what chemical should you use with optics

alcohol, such as ethanol

why is a powerful light source needed

because fluorescence has a low yield and the microscope collection efficiency is low

why must you use extreme care when cleaning with chemicals

can migrate into internal areas or could remain in contact with the surface for too long

mechanical maintenance my include

dirty body, loose nose piece, sticky stage, sticky focus / condenser / diaphragm

what results in fluorescent microscopes

fluorescing areas shines brightly against a dark background

dirt that is non adherent may be removed from less delicate surfaces by ...

gently brushing with a clean camelhair brush

what are the three main features of an vertical illuminator

heat filter, neutral density filters, and a filter cube

what is a safer alternative to xylene

histolene and histoclear

what is the basic task of a fluorescence microscope

illuminate the specimen and separate the much weaker emitted fluorescent light from the brighter excitation light and make it visible to the viewer

why is damaged cement bad?

it is irreparable and must be discarded

the darker the material, the ....

more efficient the instrument

what should you never do with cleaning a microscope

never disassemble an objective, don't clean objective back focal lens, don't apply force, and keep acids/bases away

can barrier filters filter out reflected UV light

no

can excitation filters produce fluorescence

no

what is the forensic application with fluorescent microscopy

not used to identify, only used as a comparative tool

what is fluorescence

optical phenomenon in which certain materials absorb a short wavelength, high energy photon, and re-emits it as a lower energy photon with a longer wavelength

dust, lint, and smudges on the optical components contribute to ...

overall degradation of microscope performance

example of primary fluorescence

paint or fiber

do you need to stain the samples?

possibly, some items fluoresce by itself, like scorpions

what is one danger with solvents and cleaning opticals

potential to dissolve the cements

why do we cover microscopes

prevent the microscope from being exposed to airborne contaminants protect against scratches and spills protect against damage

what is an abrasive slurry

saliva and dirt

how can you remove dust from a microscope

small vacuum, cleaner, blower, or dabbing with moist papertowel

example of secondary fluorescence

staining

all optical components should be ...

stored in some form of container to avoid getting contaminated

primary/autofluorescence

substances that show fluorescence in their natural form

secondary fluorescnce

substances that will show fluorescence only after treated with chemicals capable of fluorescing

why are excitation and barrier filters found in sets

they were designed to function together

if the background it white, the yield would be ...

to small and we wouldn't be able to see the fluorescent

if we wanted to see the emitted light in the visible region, then the excitation light has to be in the ...

ultraviolet region

the emitted light is much ___ than the excitation light

weaker (lower intensity)

what are some typical covers for microscopes

wooden, plastic, metal box, vinal, and textile

what chemical is good for removing oil and old grease

xylene

is fluorescence subjective?

yes, therefore K&Q on comparison microscope

how does a dichromatic beamsplitter mirror work

- not allowing short wavelengths through and reflects back to sample - when it comes back again, it reflects back toward the lamp - allows long wavelengths to pass though - hits barrier filter and filters again so the only thing you see is fluorescent light


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