Lecture 8

Paraffin Wax Impregnation

bakit hindi >60? shrinkage and hardening

paraffin oven 2-5

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 dapat always filtered before use

pwede 2x gamiting

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 paraffin only used 2x

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 manual or hand

ilan changes ng wax ginagamawa 4x with 15 mins interval

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 immerse in medium for approx 3 hours

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 lower MP
sliding sledge ester

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 kasi water sol = bawal dumikit sa basa kasi matutunaw
cannot be floated out on water use other solutions

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 Dry eyes ni Gilson

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 gilsons mixture sa dry pinapatak lang hanggang mamuo
sa wet celloidin nilulublob

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 double embedding = CP kasi celloidin paraffin
ICEP
Infiltrate with celloiding and Embed with Paraffin

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 1. L pieces can be adjsuted

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 anong pinagkaiba nila magkaiba size ng molecule
pinaka maliit ay spuur
medium glycerol

bisphenol largest size ng molecule

mas maliit ang size ng molecule mas mabilis mag infiltrate
spurr mas mabilis

Polyester 1950s

mas maganda epoxy

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 Trimming

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 2 kinds of trimming
coarse kapag pangit dimension ng tissue

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 Sectioning or Cutting

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 Section cut is picked by:
after cutting → transfer to floation water bath
camel or squirrel or stable

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 floation 30 seconds or less pag tumagal masisira
45 - 50

Adhesives
hindi na daw kelangan as long as chemically clean glass slide
ginagamit if not sticking to slide

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 1. xylene = milky
repeat using aOH

clearing = dealcoholization

first = fixation problem

hardening of tissue 2ndary aim
insensitive to subsquent process

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 infiltration = airholes = repeat impregnation

4-6 micra 5th paraffin celloidin 10-15 micra

6th if disposal blade = replace blade; conventional steel knives

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 to increase humidity blow or boil water to form steam??

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 terminologies
stain itself
staining result

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 mordant important sa hematoxylin

without mordant = hematoxylin not possible

examples

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 not essential compared to mordant

kahit wala may color parin

used both for indirect

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 walang differention na nagaganap
example
H and Eosin for frozen sectioning

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 exampe: routine H and E
meron decolorization

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 most common decolorizer: acid alcohol (HCl and ethyl aOH)
used for regressive staining technique using routine HE

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 kung ano color stain ganon din color

example: yellow tapos nagiging red yung tissue

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 responsible of conversion of color to blue or blue black
use weak alkali solution

tap water weak alkali

Metallic impreg
using colorless salts reduced to black deposits on surface of tissue

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 1. silver chloride

2. gold chloride

not absorbed sa surface lang

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 picro thionine for caliculi
cryptococcus = india ink - negative stain

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