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STRUCTURES - WEEK 1 > LEC 4: Visualizing Cells and Tissues: Stains - 08.18.2014 > Flashcards

LEC 4: Visualizing Cells and Tissues: Stains - 08.18.2014 Flashcards

1
Q

Hematoxylin

A
  • acts as a basic (positive) stain
  • stains deep blue-to-purple color
  • positively charged dye so binds to acidic (negative)
  • basophilic structures (structures stained display basophilia)
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2
Q

Cell structures that are basophilic (bind with hematoxylin)

A
  • DNA
  • RNA
  • Heterochromatin and euchromatin
  • nuclei
  • nucleoli
  • ribosomal particles
  • RER
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3
Q

Eosin

A
  • stains pink-red-orange color
  • acid stain (negatively charged dye)
  • binds to basic (positively charged) structures
  • structures stained display acidophilia (eosinophilia)
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4
Q

Cell structures that are acidophilic (bind with eosin)

A
  • cytoplasmic proteins
  • mitochondria
  • filaments
  • smooth ER
  • secretory proteinaceous granules
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5
Q

Under what circumstances would the cytoplasm be more blueish

A

when cytoplasm contains lots of RER (e.g. plasma cells) or free ribosomes (rRNA), cytoplasm will be more blueish

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6
Q

What is the difference between euchromatin and heterochromatin

A
  • euchromatin, DNA is “loose” for active transcription of RNA
  • in heterochromatin, DNA is condensed, inactive transcription of RNA
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7
Q

What does the nucleolus do

A
  • made up of proteins, RNA
  • transcribes and modify rRNA, integrate ribosomal proteins to form immature ribosomes
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8
Q

What does the rough endoplasmic reticulum do (RER)

A

ribosomes and rRNA making up the RER are charged with secretory protein synthesis

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9
Q

Free ribosomes

A

cytosolic protein synthesis

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10
Q

What does the smooth ER do

A
  • lipid synthesis
  • alcohol and drug metabolism
  • glycogen storage
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11
Q

What does the Golgi apparatus do

A
  • carbohydrate synthesis and secretion
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12
Q

What does the Mitochondria do

A

ATP production

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13
Q

What do microtubules do

A
  • movement of molecules, motility
  • mitosis
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14
Q

What do filaments do

A

support, contraction

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15
Q

What are the functions of lipid droplets and glycogen in the cell

A

energy sources

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16
Q

What are the major structures in this image

A

Hepatocytes (liver cells)

  • nucleus is basophilic
  • nucleolus and heterochromatin are very dark
  • euchromatin is not well stained
  • cytoplasm is pink (mitochondria, SER, filaments)
  • RER is basophilic
  • Golgi, lysosomes, glycogen, and fat are lightly stained or not stained
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17
Q

In this image of a plasma cell, what do the arrows point to

A

A. RER in cytoplasm is basophilic

B. Golgi apparatus is not stained well, appears as pale/light pink area just off nucleus (hof area)

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18
Q

Which two (2) organelles are involved in the synthesis of antibodies (glycoproteins)

A

Golgi and RER

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19
Q

histochemistry

A
  • chemical methods for detecting chemical make-up
  • identify specific molecules in cells and tissues
  • histochemical methods are also colorful (like H&E stains) but more specific
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20
Q

Feulgen Reaction (“foil-gun”)

A
  • for nuclear DNA
  • mild acid hyrolysis to remove nucleotide from deoxyribose
  • forms aldehyde group with can react with Schiff reagent (magenta color complex)
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21
Q

What are the cell phases in this garlic root

A

Feulgen reaction staining nuclear DNA

A. Interphase (nuclear membrane/envelope, euchromatin, heterochromatin, and nucleoli are present)

B. Prophase (coarse clumps of heterochromatin signify that chromatin is condensing, continues until end of prophase when nucleolus and nuclear envelope have disappeared)

C. Anaphase (chromatids separate from each other and move toward opposite poles of the mitotic spindle)

22
Q

Periodic Acid-Schiff (PAS) Reaction

A

histological reaction that detects carbohydrate-rich macromolecules

  • mucus
  • glycosaminoglycans (GAGs)
  • glycoproteins
  • glycogen
23
Q

PAS stain of intestinal epithelium

A
  • mucus of goblet cells stained deep red
  • glycoprotein of glycocalyx
  • basement membrane (heparan sulfate = GAG)

NB: These are all carbohydrate-rich macromolecules, which is why they appear in PAS stain

24
Q

What is the difference between the PAS stain of liver cells (hepatocytes) and the unstained?

A

diastase was used as a control to digest glycogen before PAS stain applied (no carbohydrates to stain)

25
Q

diastase

A

a control method; enzyme digests glycogen prior to PAS staining

26
Q

metachromasia

A

staining reaction in which the cell components are stained a different color from the dye color

27
Q

Toluidine Blue

A
  • cationic dye (positively charged)
  • metachromatic property
  • deep blue color
  • stains heparin in mast cells (highly sulfated GAG)
28
Q

H&E stain of Heparin in mast cells

vs.

Toluidine (tah-luah-deen) blue stain of Heparin in mast cells

A

Heparin in mast cells stains acidophilic with H&E (pink)

vs.

Heparin in mast cells stains magenta with toluidine (metachromatic reaction)

29
Q

Structure of Heparin

A
  • Heparin is a glycosaminoglycan (GAG)
  • repated disaccharide units
  • glucosamine and uronic acid
  • residues are sulfated, so HIGHLY NEGATIVELY charged
30
Q

Sudan IV

A

Fat stain

  • oil soluble dye with a deep red color
31
Q

Frozen section of adipose tissue, what stain is used

A

Sudan IV stains fat in adipocytes

32
Q

Osmium tetroxide (OsO4) Stain

A

Lipid Stain, strong oxidizing compound

33
Q

In this slide of myelinated axons, what stain is being used

A

OsO4 is staining the fatty (lipid) myelin of the axons black.

  • Note Node of Ranvier where myelin sheath is not present
34
Q

Resorcin-fuchsin Stain (re-source-in fuuksin)

A

stains elastic fibers in extracellular matrix black

35
Q

H&E Stain of elastic fibers

vs.

Resorcin-fuchsin Stain of elastic fibers

A
  • H&E Stain, elastic fibers are acidophilic (pink)
  • Resorcin-fuchsin Stain, elastic fibers stain black
36
Q

Mallory Trichrome Stain

A
  • more sensitive than H&E
  • routinely used in clinics for histopathological diagnosis of disease related to fibrosis
    • hepatic fibrosis
    • pulmonary fibrosis

NB: fibrosis = formation of excess fibrous connective tissue in organ/tissue in a reparative or reactive process

  • hepatic fibrosis = excessive connective tissue builds up in liver (can lead to hypertension, scarring distorts liver blood flow, or cirrhosis)
  • pulmonary fibrosis = excessive connective tissue in lungs hinders lung function (shortness of breath)
37
Q

H&E Stain of collagen fibers in connective tissue

vs.

Mallory Trichrome Stain of collage fibers (Type I and III) in connective tissue

A
  • collagen fibers are acidophilic (pink) in H&E Stain
  • collagen fibers (Type I and III) stained blue in Mallory Trichrome Stain
38
Q

Silver stain

A

Stains Type III collagen (a.k.a. reticular fibers) black

39
Q

Stain of Type III collagen (reticular fibers) in spleen

A

reticular fibers are stained black with silver stain

40
Q

enzyme histochemistry

A

permits localization of enzyme activity in a tissue section using:

  1. substrate of enzyme
  2. capturing agent (will combine with enzyme product to form insoluble precipitate, visible)
41
Q

Acid phosphatase

A

marker enzyme for lysosomes

42
Q

Azo dye method

A
  • Uses diazonium as capturing reagent for acid phosphatase
  • carried out at pH=5
  • produces a Diazonium complex that appears RED
43
Q

Kidney tubule cells treated with Azo dye method

A
  • Azo dye method produces a Diazonium that appears RED
  • localizes acid phosphatase
  • presence of lysosomes
44
Q

Succinate dehydrogenase (SDH)

A

oxidate enzyme of the citric acid cycle (TCA), localized in mitochondria (marker of mitochondria)

45
Q

Tetrazolium method

A
  • uses Tetrazolium as capturing reagent
  • produces insoluable black Formazan under physiological temperature and pH
46
Q

Submandibular gland treated with Tetrazolium method

A
  • SDH found in ducts (black appearance)
  • indicates presence of mitochondria
47
Q

immunohistochemistry

A

permits detection of proteins in cells or tissue sections based on reaction between antigen and its antibody

48
Q

What are the two (2) types of immunohistochemical methods

A
  1. antibody marked with fluorescent compound
  2. antibody marked with peroxidase
49
Q

Diaminobenzidine (DAB) method

A
  • demonstrates peroxidase activity
  • creates insoluble brown color compound
  • DAB oxidized to insoluble brown compound at site of peroxidase activity
50
Q

Actin filaments (2 different treatments)

A

L - detection of actin filaments with actin antibody (antibody for actin protein) labeled with fluorescent dye

R - detection of actin filaments with actin antibody labeled with peroxidase (DAB method)

NB: DAB method oxidized to insoluble brown compound at site of peroxidase activity

51
Q

Immunoperoxidase staining for insulin in islet of Langerhans of pancreas

A
  • insulin is a peptide hormone
  • antibodies for insulin tagged with peroxidase
  • DAB (diaminobenzidine) oxidized to insoluble compound at site of peroxidase activity (brown color)

STRUCTURES - WEEK 1 (8 decks)

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