PART 2 - HP - IHC, CYTO fr Sir Ed Flashcards
guidelines for sputum collection?
-Obtain at least 3 consecutive morning specimens through deep cough
-Inhalation of aerosol solution for 20 minutes can help induce coughing of sputum
NOT—> Collect sputum using a wide-mouthed jar with formaldehyde fixative
Wound from friction or rubbing against a rough surface:
Abrasion (open wound)
Cut or tear in the skin from a sharp object:
Laceration (open wound)
Caused by a sharp, pointed object that penetrates the skin:
: Puncture wound (open wound)
Blood collection outside blood vessels due to trauma:
Hematoma (closed wound)
Bruise from blood leakage into tissues after trauma:
: Contusion (closed wound)
IHC
Animal used for production of monoclonal antibodies:
Mouse
IHC
Polyclonal antibodies:
Rabbit, goat, pig, sheep, horse, guinea pig
IHC
The first step in immunohistochemistry:
FIXATION
IHC
A preparation step prior to staining that only applies to immunohistochemistry:
Antigen retrieval
IHC
Antigens are most likely to be demonstrated at max sensitivity with:
: Frozen sections fixed in acetone
**Preparing tissue for immunohistochemistry: In certain instances, the tissue must be prepared as a cryostat
section and fixed for a few seconds in absolute methanol or acetone, to preserve immunological activity and prevent destruction of some of the labile antigenic sites. However, immunofluorescence and immunoperoxidase techniques may also be done on formaldehyde-fixed and paraffin embedded sections.
IHC labels?
Enzymes (HRP, ALP)
microscope used for IHC?
Bright field microscopy
labels for Immunofluorescence (IF) ?
✓ Labels: Fluorescent dyes (FITC, TRITC)
microscope used for Immunofluorescence (IF) ?
Visualization: Fluorescence or confocal microscopy
In IHC, many masked antigens can now be retrieved in routinely processed tissue by:
✓ Proteolytic enzyme digestion
✓ Microwave antigen retrieval
✓ Microwave
✓ Trypsin antigen retrieval
✓ Pressure cooker antigen retrieval
Horseradish peroxidase (HRP) and alkaline phosphatase (ALP), which convert 3,3’ diaminobenzidine (DAB) and 3-amino-9-ethylcarbazole (AEC), into _____ and _____ end products, respectively
Brown and red
A relatively new technique that involves the boiling of formalin-fixed deparaffinized sections in certain solutions, such as 0.01 M Citrate/Citric acid buffer (pH 6.0), EDTA (pH 8.0) or Tris EDTA (pH 10.0): M
Microwave antigen retrieval
Used in rapid fixation, rapid embedding, special staining and antigen retrieval in IHC:
Microwave processing
it works as a physical agent to increase the movement of molecules and accelerate fixation.
Microwave
✓ Also used to accelerate staining, decalcification, IHC and EM
✓ Major advantage: tissue is heated right through the block in a very short time
microwave
Antigenic determinants masked by formalin-fixation and paraffin embedding often may be exposed by epitope unmasking, enzymatic digestion or saponin, etc. Do not use this pretreatment with frozen sections or cultured cells that are not paraffin-embedded.
PRE-TREATMENT OF TISSUE SECTIONS
referred to as Cancers, derived from “cancrum”, a Latin word for crabs
Malignant Neoplasms:
Malignant tumor of Connective tissue (Bone, cartilage, fat, muscle, blood vessels or other connective or supportive tissue)
Sarcoma
Malignant tumor of Epithelial tissue (skin or tissues around internal organs)
Carcinoma
✓ Benign tumor of Epithelial tissue
✓ Derived from the ducts and acini of glands, although the name is also used to cover simple tumors arising in solid epithelial organs
Adenoma
✓ Benign tumor of Epithelial tissue
✓ Take origin from an epithelial surface
Papilloma
Benign tumor of the Striated muscle
Leiomyoma
Benign tumor of the Smooth muscle (heart muscles)
Rhabdomyoma
Malignant tumor of the Muscle
Myosarcoma
Malignant tumor of the Striated muscle (skeletal muscles)
Rhabdomyosarcoma
Malignant tumor of the Smooth muscle
Leiomyosarcoma
type of cell comprises ducts of glands and walls of thyroid follicles?
simple cuboidal
Nerves cannot be donated due to?
complex neural connections, risk of neurological damage and limited regeneration capacity
it is the outermost layer of an organ, a thin membrane surrounding abdominal organs (e.g., intestine, stomach), thoracic organs (e.g., heart, lungs) and reproductive organs (e.g., ovaries, testes)
Serosa
it is the outer layer of an organ with specific functional and structural roles, as found in the adrenal gland, kidney, and brain
cortex
What would be the appearance of superficial cells in Papanicolaou stain?
Orange to pink
it deals with the microscopic study of cells that have been desquamated from epithelial cells
exfoliative cytology
it is useful for lesions found in organs such as breast, thyroid, and lymph nodes.
Fine needle aspiration
CEA can be used to differentiate adenocarcinoma from mesothelioma. Between the two, which is positive for CEA?
adenocarcinoma
What additional step is done in IHC for paraffin- embedded tissues?
Antigen retrieval
What is the use of serum blocking in IHC?
To avoid unwanted non-specific staining.
Primary antibody is directly conjugated with a label, what IHC technique is this?
Direct IHC
IHC technique which involves the use of a bridging antibody that links the soluble peroxidase-antiperoxidase complex to the unconjugated primary antibody.
PAP technique
One of the commonly used chromogen in IHC that gives dark-brown end products.
DAB
