السلام عليكم ورحمة الله

ساقوم بوضع مبادئ للعملي المايكرو وان شاء الله تستفيدوا منه
اولا

staining technique

***stain***
chemically when dissolve give ions
**aims of staining**
_study morphology of bacteria(cocci_bacilli_spirilla_vibrios)

study arrangment
فمثلا ممكن تكون في شكل single مثل escherichia coli

_diplio or pairs مثل klebsiela rhinoscleromatis و streptococcus pyogenes
او في شكل عناقيد( cluster) مثل streptococcus aureus( المكورات العنقودية الذهبية)
او ممكن تكون البكتريا في شكل حلقات (chains) مثل streptococcus pyogenes
_او متفرعة (branched) مثل nocardia africana


_(to study special structure (organelles
مثل capsule و flagella و spore
_classification of bacteria into gram+ve و gram_ve


**classification of stains**
*according to
:sm187:radicle which carry dye (colour
*acidic stain*
colour contained in acidic part while basic part colourless
e.g eosin stain
*basic stain*
colour contained in basic part while acidic part colourless
_major of stain in microbiology lab ar basic
e.g *crystal violet in gram stain
*methylene blue
*basic fuchsin in z.n stain
*neutral stain*
colored contained on acidic and basic parts
e.g giemsa stain
التقسيم على حسب طبيعة الصبغة*** according to nature (source) of stain
*natural stain*
e.g carmine stain, haematoxyline
*synthetic stain*
ستخدم alanine compound
e.g gram stain و loefflers methylene blue
التقسيم على حسب عدد الصبغات** number of stains***
*simple stain ومثال عليها loefflers methylene blue وهي التي يستخدم فيها نوع واحد من الصبغات
*compound stain*
contain 2 or more than one stains
ومثال عليها gram stainالي تحتوي على crystal violet كصبغة اساسية basic stain) ,و safranin كاونتر (counter stain)
* التقسيم على حسب التفاعل
*according to reaction of stain
قسمت الى
*direct stain
stain bacterial cell directly without need aid of other chemical substances
ومثال عليها loefflers methylene blue
*indirect stain
stain bacterial cell with need aid of chemical substance
وهذه المواد الكيميائية هي
*mordant*
chemical substance addae before or during staining for complete staining reaction and linking between cell and stain
ومثال عليها
lugols iodine in gram stain
phenol in z.n stain
*accelerator*
chemical substance used to speed reaction of stain and make reaction more selective
e.g
KOH(potassium hydroxide) in loefflers methylene blue
*decolorizer*
chemical substance used for removal excess of stain and differentiation of bacteria into gram_ve and gram+ve in gram stain

مثال على decolorizer
70% alcohol in gram stain
و
acid alcohol in z.n stain






***********************************************
اول نوع من الصبغات هو
loeffler"s methylene blue

Difinition:sm198::sm188:
simple direct rapid and basic stain
AIMS
study morphology of bacteria
study arragment of bacteria
study colour of bacteria
also use to
stain leucocyte in faecal specimen
detect bipolar morphology


Requirements:sm189::sm189:
clean dry slide
wire loop
isolate (pure and fresh(
sterile normal saline
bunzen burner(for sterilization of loop and fixation of smear)
loefflers methylene blue stain
microscope (x100)
oil

STEPS
prepare smear(small drop of normal saline +portion just touch of inoculum)
mix well then spread at circular manner
air drying
fixation by gentel heat
cover the smear with loefflers methylene blue for 1_2 minutes
wash the smear
wipe the background( clean)
place on drop of oil
examine under microscope (x100)
record your result

resulr:sm178::sm178:
morphology: cocci_ bacilli
arrangment: cluster_ single
colour: blue

: