Enzyme Histochemistry & Electron Microscopy

The Sxns of muscle stained with the NADH procedure show the presence of marked participate. The most likely cause of this is

a ph above 8

Microscopic examination of snxs stained with napthol-AS-D chloroacetate esterace technique shows refractile areas in nuclei, and a brown, granular, pigment like artifiact scattered throughout the sxn. This was most likely caused by

Not dipping in xylene before mounting. (its called cornflaking)

The final colored product in the ATPase reaction (dark red-bown/ lighty colored to unstained)

Cobalt Sulfide

Micropscopic examination of sxns of skeletal muscle stained with the a-napthyl acetate esterase stain reveal a yellow precipitate on sxns. This could most likely have been prevented by:

washing sxn more thoroughly after staining

Some fibers in this image are stained black bc they are (a-napthyl acetate esterase stain)

denervated. the dark fibers are smaller and darker with the a-napthyl acetate esterase stain bc they are denervated & a disease has occurred.

What is the best long term storage procedure for storing frozen muscle tissue used for enzyme histochemical stain

at -70C to -80C

which enzyme activity is histochemically detectable in greater amounts in slow (Type 1) fibers of skeletal muscle?

succinic dehydrogenase

Slow muscle fibers which use oxidative metabolism to generate ATP contain

high levels of oxidative enzymes.

An enzyme is a

protein

Skeletlal muscle fibers can be identified by cross striations and

peripheral nuclei

This ATPase stain was done at 4.3. the very dark fibers are

type 1. when the ATPase stain is done at 4.3 the type 1 fibers will stain dark and the type 2 fibers will stain dark.

which component of the incubation medium used in the esterase method for the demonstration of neutrophils is the substrate?

naphthol AS-D chloroacetate`

which of the following enzyme methods is most frequently used to demonstrate the different muscle fiber types

ATPase or adenosine triosphatase

Lysosomes look like____ under EM

Round black circles

All enzymes:

change the rate of chemical reactions

which of the following chemical reactions from the final visible reaction product in the calcium cobalt method for ATPase

cobalt phosphate + sulfide ions = cobalt sulfide

the primary problem in this tissue (blue, Swiss cheese/ hole-y looking thing)

poorly infiltrated

tetrazolium salts such as nitro blue tetrazolium

change to a colored product when reduced

the endoplasmic reticulum will increase

basophilic staining, or uptake of hematoxylin

several of the frozen sxns of muscle tissue have been fixed in acetone. which of the following techniques would possibly show positive results with these slides?

phosphorylase

the material labeled 1 in the image is known as

euchromatin. will not stain with hematoxylin

electron micrographs of a kidney biopsy show electron Lucent membranes. this is possibly the result of

no fixation with osmium tetroxide

the a-naphthyl acetate esterase stain will stain motor endplates:

brick red and normal muscle fibers pale yellow

the histochemical study of adenosine tropsphatase (AtPase) activity in skeletal muscle biopsies require:

no fixation, cryostat, then no post fixation of cut sxns

which of the following techniques can be used to show different types of white blood cells

chloroacetate esterase

in the image which number is most responsible for the uptake of hematoxylin?

2

the small wavy structure identified by the arrow in the EM image is

Golgi apparatus, will show negative or no staining

The type of knife used most frequently to cut ultra thin sections for electron microscopy is:

diamond

for the atpase stain the ph meter should be calibrated between which of the following

4.0-10.0

if no enzyme activity is detected on histochemically stained sxns of skeletal muscle then the tech should repeat the stain:

also staining a sxns of tissue known to contain the enzyme

adequate fixation of a kidney biopsy specimen for diagnostic EM is obtainable with all of the following with the exception of initial fixation in:

95% alcohol followed by glutaraldehyde

the best medium for mounting muscle tissu for freezing is

gum tragacanth

the stains on a muscle case were not read for three days after completing and the phosphorylase stains are totally negative. what is the quickest way to determine whether r not the sxns have faded?

remove the coverslip and retain in dilute gram iodine

SDH stains

a blueish violet color

enzymes that act on substrates by removing water are classified as:

hydrolases

In order to ensure stable sections in the electron beam, tissue for electron microscopy is embedded in:

epoxy resin

when adjusting the ph of a solution for enzyme histochemical staining:

adding sodium hydroxide will make the ph higher

the irregular, dense, structures seen in the capillary loop of this renal biopsy are

immune complexes

biological reduction occurs with the loss of which of the following

oxygen

the instrument used to cute thin sxns of epoxy resin embedded tissue for electron microscopy is called an

ultramicrotome

image shows muscle that is pink with peripheral nuclei surrounding. what type of muscle is this

skeletal

when the primary fixation of tissue for EM is done with aldehydes fixatives then the secondary fixation is usually done with

osmium tetroxide

the arrows shown in the image are: ( looks like circles but made with lines, they are not lysosomes or mitochondria)

centrioles

image shows pink muscle with ice crystals that looks like "holes or Swiss cheese" what caused the artifact

slow freezing

A small tissue sample is important for electron microscopy because:

osmium tetroxide penetrates very poorly

microscopic examination of an atpase stain seen of skeletal muscle reveals muscle fibers appearing as thick rimmed circles with empty centers. this is a sign of

slow freezing