Exam 1 9/13 Howard Intro to Histology Flashcards
Another name for histology
Microanatomy
Methods for viewing tissue
Conventional Light Microscopy
Confocal light microscopy
Transmission Electron microscopy
Scanning electron microscopy
X-ray
Computer Axial Tomography
Magnetic resonance imaging
Positron emission tomography
Which method of viewing tissue will we normally see/sometimes see?
Conventional light microscopy; transmission electron microscopy
True or false: confocal microscope has only one lens
False - more than a single lens
What magnifications are possible with a compound light microsope?
40, 100, 400, 1000x
True or false: electron microscopes can provide a 2D and 3D image
True
Transmission electron microscope provides what kind of image?
2D
Scanning electron microscope provides what kind of image?
3D
True or false: you cannot see small molecules with an electron microscope
False - can see up to 1nm which is the size of some small molecules, like glucose
Approximate size of cell nucleus
1-10 micrometers
Resolution of light microscope
200 nm
When viewing artifacts, what is your ruler?
identify different cells and nucleus as a reference point
What affects the appearance of artifacts?
- Preservation/fixation
- embedding
- sectioning
- staining
What is usually used in fixation of a specimen?
37% buffered formaldehyde
What is usually used to embed a specimen?
Paraffin wax
What is usually used to stain a specimen?
H&E (hemotoxylin and eosin)
True or false: when preserving a specimen, they are hydrated, dehydrated, then rehydrated again
False - first dehydrated, rehydrated, then dehydrated to apply stain
True or false: thinner section allow for better resolution
True
Embedding methods
- frozen
- paraffin
- plastic
Pros and cons of frozen specimen
Pros:
1. quick to retrieve, cut, stain
2. can preserve antibody-antigen complexes
Cons:
1. can destroy antibody-antigen complexes
2. morphologically destructive
3. poor resolution
Pros and cons of embedding specimen in paraffin wax
Pros:
1. very standardized
2. standardized interpretation
3. mostly preserve antibody-antigen complexes
Cons:
1. long fixation times
2. resolution can vary with technician
Pros and cons of embedding specimen in plastic
Pros:
1. very good resolution
2. water soluble plastic retains lipids
Cons:
1. long fixation times
2. interpretation can vary depending on procedure
3. destroys antibody-antigen complexes
What is used to section specimen?
microtome, cryomicrotome
How thin should a frozen section be?
5-100 micrometers
How thin should paraffin sections be?
2-50 micrometers
Plastic for light microscopy section size
0.5-2 micrometers
Plastic for electron microscopy section size
40nm - 100nm
3 classes of staining:
- differentiate between acidic and basic components of cell
- differentiate fibrous components of ECM
- metallic salts that precipitate metal deposits on tissues
Acid stain
Eosin; negatively charged dye; stains basic structures in cytoplasm and mitochondria, structures are acidophilic
Basic stain
Hematoxylin; positively charged dye; stain acidic structures (DNA and RNA); these structures are basophilic
What color does hematoxylin stain?
Blue (dna and rna)
What color does eosin stain?
Pink (Basic proteins in cytoplasm - collagen, elastin, reticular fibers, but can’t distinguish)
What does H&E not stain?
Lipids - they are lost in preparation, as they are soluble in dehydration reaction alcohols
Silver stain can stain:
reticular fibers black (type III collagen)
Weigert’s elastic stain
Blue - elastic fibers
Periodic acid-schiff
Magenta - glycogen and carbohydrate rich molecules
4 Primary tissue types
- Epithelial
- Connective
- Muscle
- Nervous
Epithelial tissue are specialized to:
protect, absorb, and secrete
Epithelial tissue function
- covers and lines (skin, lining of gut)
- modified for absorption (gut lining)
- modified for secretion (glands)
Connective tissue is specialized for:
providing support and holding other tissues together
What connective tissues are specialized?
blood/bone marrow, mineralized tissues (ex: teeth), fat storage
______ tissue is highly specialized for contraction and conduction
Muscle
____ tissue is highly specialized with respect to irritability and conductivity
Nervous