Histo

*Specimen that should be completely dried befor fixing: blood

*Best fixative for cytologic smears: 95% ethanol
*Site of collxn for dyslplasias and carcinomas of the cervix: T-Zone
*gynecologic specimen should include all the : squamous, columnar & metaplastic cells
*Sputum: at least 3 consecutive morning and 2-4 slides(1for Giemsa and 2 for Pap’s)
*Lateral vaginal scrape: for hormonal evaluation
*concentration method for pleaural fluid and/or other fluids in small quantity: cytospin
*Papanicolau(Pap) smear: staining method for exfoliative cytology
*saccomno preservative: 50% alcohol and 2%Carbowax (answer: Carbowax)
*method for palpable masses: FNAB
*all are exfoliative except: thyroid follicles
*Paps smear: blue cytoplasm
*ammonium hydroxide: bluing agent in paps
*minimum amount for pleural fluid: 20-30ml (30ml ang nasa choices)
*minimum fixation time for paps smear: 30mins
*progesterone-estrogen effect: Navicular cells
*needle gauge for FNAB: 22-23 nasa grego(magkaibang choices pero nasearch ko sa net
e 23 ata? Pacheck guys)
*wright Giemsa is only used for: CSF
*Mature superficial cells: polyglonal squamous cells, pale pink cytoplasm,pyknotic
nuclei, true acidophilia(influenced by Estrogen)
*Endometrial cells: shed in response to ovarian hormones, 1-10 days after menstruation
*endocervical cells: honeycomb appearance
*maturation index: superficial cells, intermediate cells ,(and parabasal cells not so sure)
*confirmation for Sputum: alveolar macrophage
*abstinence for gynecologic specimen: 1 week
*haparin: AC of choice. no interference in cytologic preparation & eval’n
*apparatus used to collect vaginal secretions ecept: ayre’s spatula, brush, speculum,
answer: pipet & rubber bulb
*anticoagulant for body fluids: heparin

Headings:
~~~~Types of knives
*plane concave:
Less concave-celloidin-embedded on a sliding microtome
More concave-paraffin sections on base-sledge, rotary or rocking microtome
*biconcave:paraffin embedded on a rotary microtome
*plane wedge knife: for frozen sections,paraffin embedded using base-sledge or slidingM
~~~~Washing-out: tap water, alcohol iodine, 50-70%alcohol
*Tap water: removes chromates, osmic acid , formalin “COF”
*50-70% alcohol: removes picric acid
*alcoholic iodine: removes mercuric fixatives

~~~~Blocking out molds:

*Leuckhart’s Embedding mold
-L-shaped strips
-flat metal plate w/c can be moved to adjust mold to the size of specimen
-for routing use
-cumbersome for bc labs
*Compound Embedding Unit:
-interlocking plates resting on a flat metal base
-with several compartments
-adv:embeds more specimen at a time so it reduces time
*Plastic Embedding Rings and Base Mold
-special stainless steel w/ embedding ring
*Tissue Tek
-warm plate and cold plate
-adv:ease of use,less oaraffn wax needed,firmly attchd to tissue n holder

~~~Fixatives: picric, osmium tetroxide, helly, carnoy’s, glutaraldehyde d sure if meron sa

choices
*Picric acid
-1%, yellow color, preserves Glycogen, allows brilliant staining w/ Trichrome Mx
-a component in Bouin’s Sol’n fixative for Embryos
-explosive when dry
*Carnoy’s Fluid
-for urgent biopsies so most rapid fixative
-preserves Nissl granules, cytoplasmic granules
*Osmium tetroxide
-pale yellow powder
-strong oxidizing sol’n
-fixes fats & lipids and r stained black
*acetone: -5 to 4C

Xylene, paraffin, acetone,

*preservative in egg albumin: thymol

*temperature of flotation bath: 45-50C
*paraffinoven: 55-60C
*microwave principle: convection
*several compartments: compound
*tissue tek: warm and cold plate
*plane concave: less concave-celloidin
*biconcave
*plane wedge
*lungs: 1-2cm
*fixative for hard tissues: 4% phenol
*Stat for tissue processing :spurr
*pawl: rotates rachet feed and micrometer screw
*rachet feed wheel: regulates the thickness of the section
*for badly nicked knives: fine carborundum
*40-120: strokes of Stropping
*mineral oil: not used for Stropping
*bluing agent: lithium carbonate & ammonium hydroxide
*sections are squashed: bevel of knife is lost due to incorrect sharpening
*sections of unequal thickness are produced: blocks are too large
*used for electron microscopy: acrolein(glutaraldehyde and formaldehyde) and
Karnovsky
*fixation ratio: 20:1 except osmium tetroxide(5:1)
*decalcification: 20:1
*dehydration: 10:1
*heidenhain’s susa: type of mercuric chloride fixatives that don’t produce black pigments
*formaldehyde: white ppts
*fixative that imparts white ppts at low temps: formalin
*resinous media: canada balsam,DPX,XAM,CLARITE memorize refractive index
*aquaeous media: water,glycerin,farrant’smedium,apathy’s, brun’s fluid
*in regressive staining what is decolorized cytoplasm or nucleus?
* blue cytoplasm except: H&E(pale pink)
*dezenkerization: mercuric chloride
*nuclear fixative: flemmings with glacial
*involves mordant and accentuator: INDIRECT STAINING
*Formalin concentration in routine fixation: 10%
*temperature of Acetoone: -5 to 4C
*non additive fixative: acetic acid
*cryostat: 4 micra
*alum: ehrlich’s and Harris hematoxylin and Mayer’s
*not more than 4mm: size of tissue in fixation
*ultrathin: 0.5 micrA
*impregnating for stat: Spurr
*flywheel: for manipulation and operation
*fatty samples: freezing microtome
*site of collection for 58y/o woman:
*pale pink staining cytoplasm: Normal
*mountant of Cryostat OCT, water, bovine albumin, von apathy gum
*explosive impregnating agent: LVN
*adhesive that impart blue color on the slide: albumin
*disadvantage when using high concentration of formalin: over hardening
*preservative added to glycerin: thymol
*Impregnation or infiltration: process where in clearing agent is removed replacing a medium that fills
the tissue cavities
*alcohol fixatives causes: Polarization

NICE TO KNOW: 
*Osmium tetroxide: fixative for EM, it contains Sucrose
*common concentration: 10% Formaldehyde, 3% Glutaraldehyde
*0.25% Glutaraldehyde: for ImmunoEM
*penetration of fixative: 1mm per hr.
*osmium tetroxide: inhibits hematoxylin staining
*fixative for Lipids: mercuric chloride & potassium dichromate
*for Carbohydrates: alcoholic fixatives
*mixture fixatives: Karnovsky’s, acrolein
*removal of white ppts in formalin and to prevent decomposition : 10% Methanol
* bleaching of fixative is prevented by changing fluid every: 3 MONTHS
*cadmium & cobalt: to prevent dispersion of fats into fixative
*fixation by formalinin influenced by: HEAT, VACCUM, AGITATION, MICROVAWE TECHS.
*fixation is retarted by: large n thick tissue, Mucus, Fat, Blood, Cold Temp
*shrinkage and swelling of cells: OVERFIXATION
*prevent “gratings”: use 10% Hydrochloric Acid as decalcifying agent
*5-10% Nitric Acid: most common and fastest decalcifying agent
*Parenyi’s Fluid: softens and decalcifies
*Hydrochloric acid: for surface decalcification
*chelating agents:
EDTA(versene)-binds Ca & Mg, for detailed microscopic studies
Ion Exchange Resin: not for fluids w/ mineral acids, nitric acid, HCl
*best to measure degree of decalcification: physical or X-ray Mx
*decalcifying fluid is changed evry: 24-48hrs or 1-2 days
*remove decalcifying agent with: NSS
*tissue softeners: Molliflex, 2% HCl, 1% HCl w/ 70% alcohol
*dehydration:
-removes water following fixation
-increasing strength of alcohol
-70%—95%—100%
*for embyonic tissues: 30% ethanol
*ethyl alcohol: for routine
*Methyl alcohol: for blood and tissue films
*acetone: cheap and rapid acting, extme volatile n highly flammble
* Dioxane: Dehydrating and clearing, expensive n explosive
*Celloslove: ethylene glycol monoethyl ether
*tetrahydrofuran: dehydrates and clears, causes conjunctival irritation
*clearing: de-alcoholization, removes dehydrating agent and replace a substances that will dissolve the
wax or the medium on w/c the tissue is to be mounted(eg.Canada Balsam)
*xylene: most common clearing agent
*glycerin and gum syrup:if tissue is cleared w/ water, de-alcoholization is omitted
*xylene becomes milky: incompelete dehydration
*benzene: causes aplastic anemia
*chloroform: toxic to liver, up to 1 cm thick of tissue for tough tissues
*cedarwood oil: clear both paraffin and celloidin sections, for CNS tissues
*Aniline oil: insect and delicate specimens
*Clove oil: it has tendency to be adultered
*Embedding/ Blocking/ Casting
*paraffin wax melting point: 58C
*wax bath: should be 3C above the melting point of the wax
*automatic processing: autotechnicon ex. Elliot bench-type processsor
*Vaccum Embedding: reduced from 25-75% of the normal time