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blood specimens are 3 types
1- whole blood
2- plasma
3- serum
whole blood includes ?
anticoagulant added ?
includes all cellular components (RBCs, WBCs, platelets) and fluid components
yes, it includes anticoagulant
plasma is
anticoagulant ?
the liquid portion of blood without blood cells
yes it includes anticoagulant
serum is
anticoagulant ?
the liquid portion of blood (plasma) without the clotting factors
no its without anticoagulant
what is this specimen ?
does it includes anticoagulant
whole blood
yes it includes anti coagulant
what is this specimen ?
does it includes anticoagulant
blood plasma sapecimen
it includes anticoagulant
what is this specimen ?
does it includes anticoagulant
this is serum specimen
no it doesnt include anticoagulant
2 types of collecting tubes
without anticoagulant
with coagulant
3 types of blood sample
capillary
arterial
venous
when is capillary blood simple is required
required when only few drops are needed
when is arterial blood simple is required
used to measure arterial blood gases like oxygen , co2 , Ph
when is the venuous blood sample is required
its the specimen of choice for most routine laboratory
3 types if venipuncture methods
needle and syringe
butterfly needle
evacuation tube system
2 venipuncture sites
1- the cubital fossa in 3 veins (basilic veins, median cubital vein,,cephalic vein )
2-wrist and dorsum of the hands are also commonly used
9 things needed for needle and syringe venipuncture
gloves
syringes
needles
gauze sponges
tourniquet
evacuated collecting tubes
alcohol wipes
bandages
sharp container (safety box)
sample transport is characterized by 4
1. Rapid delivery to the lab
2. Seal the tube cap & Pack the specimen in an insulated, leakproof container
3. Specimens that must be cooled (e.g., ammonia) in crushed ice. 4. Wrap specimens that require protection from light (e.g., bilirubin)
what specimen must be cooled
ammonia samples
what samples requires being wrapped to be protected from light
bilirubin sample
repeating thaw and freezing what should i do
nooooooooooooooo
this is unacceptable
tips for specimen storage
1. Store specimens in an appropriate temperature
2. Repeat thaw and freezing of samples is unacceptable
3. Keep all specimen away from bright light
mention a qualitative
method
semi-quantitative
quantitative method
for glucose measurement in urine
fehling sol.
urine strip
colorimetry
principle of benedict and fehling test veeeeeeeeeeeeeeeeeery important and discuss it below
Glucose + 2Cu⁺² +2H₂O → Gluconic acid + Cu₂O +4H⁺
reducing sugars (have free aldehyde or ketone functional groups) can reduce cupric ions Cu⁺² (blue) in benedict reagent to cuprous ions Cu⁺ (RED)
benedict/fehling test disadvantage
not specific for glucose
what substance is highl specific for glucose used in its measurement
glucose oxidase
glucose oxidase method reaction is ?
its a double sequential enzymatic reaction in the presence of an indicator that changes in color in the presence of glucose
active reagents in urine glucose dipstick 2?
glucose oxidase & peroxidase
when does glucose appear in urine
glucose appears in urine when blood glucose levels are more than 180mg/dl
what is the renal threshold for glucose
when glucose level in the blood is more than 180mg/dl
what are the 2 catigories of causes of glucosuria
hyper glycemic glucosuria
normoglycemic glucosuria
hyperglycemic glucosuria is sub classified into 3
diabetes mellitus
increased anti insulin hormones
stress glucosuria
what are anti insulin hormones
thyroxine , growth hormone , adrenaline
normoglycemic glucosuria is classified into 2 types
renal glucosuria due to kidney disease
glucosuria in pregnancy (low renal threshold)
Low renal threshold معناها انه مش شرط السكر يبدا يظهر في الurine عند
180mg/dl
لا بيظهر قبلها كمان
what is the principle of protein identification in a sample using the biuret method
in the reaction cupric (Cu+2) complexes with N atoms involved in peptide bonds of protein
so violet color appears and its intensity is proportional to the number of peptide bonds which reflects protein concentration
من الاخر ال Cu+2
بيمسك في النيتروجين اللي في الامينو اسيدس الاحماض الامينيه
ولما بيتركبو علي بعض بيعمل لون بنفسجي
what are plasma proteins
they are proteins present in the blood plasma
normal range for total plasma proteins
6-8gm/ dl
types of plasma proteins include 3
1-albumin (50%-60%) normal range of plasma albumin is (3.5-5g/dl)
2-globulin (alpha,beta,gamma)
3-fibrinogen
how are plasma proteins measured
they are measured by colorimeter
how does colorimeter work
عشاان تفهمه بس
it measures the absorption of light
how does colorimeter work while measuring plasma proteins
the colorimeter measures the absorption of light so the intensity of the violet color is proportional to the concentration of total plasma proteins
albumin in urine relation to kidney health
• A healthy kidney doesn’t let albumin pass into the urine.
• A damaged kidney lets some albumin pass into the urine.
is there any protein in urine for a normal healthy individual
normally small amounts of protein are present in urine
amount of total urinary protein and total albumin in urine
normally total urinary protein excretion is less than 150mg per day
and less than 20mg/day of albumin
what does abnormal albumin levels in urine reveal
albumin in urine can reveal a developing renal pathology at an early stage
protein in urine can be detected by 4
1-urinary test strips
2-the biuret test
3-heat coagulation test
4-colorimeter and immunochemical method
protein in urine other name ?
scientific name يعني
proteinuria
active reagent in urine strips (dipsticks) for protein
tetra-bromophenol blue
فاكر بتاع ال
agarose gel electrophorises هو هو
what does urine strip color change to when proteins are present in urine
greeeeeeeeeeeeeen
urine strip sensitivity to albumin and globulins
urine stripes are highly senitive to albumin and less sensitive to globulins
lipoprotein structure formed from 2 parts
lipid part which contains 3
phospholipid
cholesterol
and triglycerids
non lipid part which contains apoprotein
2 methods for separation of plasma lipoproteins
1-electrophoresis
2-ultracentrifugation
migration of lipoproteins in an electric field depends on 2?
and we are talking about which method of separation of plasma lipoproteins ?
1- amount of -ve charge
2- molecular weight
electrophoresis
separation of plasma lipoproteins using ultracentrifugation depends on ?
ratio between protein and lipid (density)
ترتيب ال Lipoproteins
after separation using ultracentrifuugation
from up to down
chylomicron
vldl
ldl
hdl
albumins and FA
ترتيب ال Lipoproteins
after separation using electrophoresis
from cathode to anode
chylomicron
ldl
vldl
hdl
الاختلاف هنا بين ال
ldl و vldl
functional plasma enzymes
conc in plasma relative to tissues ?
function known ?
substrates presence ?
site of synthesis?
effect of disease ?
examples ?
higher conc in plasma than tissues
have known functions
their substrates are always present in blood
liver
decrease in liver disease
ex: clotting factors (prothrombin)
lipoprotein lipase
pseudo choline esterase
non functional plasma enzymes
conc in plasma relative to tissues ?
function known ?
substrates presence ?
site of synthesis?
effect of disease ?
examples ?
present in plasma in v.low conc in comparison to tissues
no known functions
their substrates are absent from the blood
different organs like heart brain liver and skeletal muscles
different enzymes increase in different organ diseases
ex: amylase & lipase
CK&LDH
ALT&AST
alkaline phosphatase & acid phosphatase
sources of non-functional plasma enzymes 4
1-increase the rate of enzyme synthesis
2-obstruction of normal pathway
3-increase permeability of cell membrane
4-cell damage with the release of its enzyme content
say an example on each of the following sources of non func plasma enzymes
1-increase the rate of enzyme synthesis
2-obstruction of normal pathway
3-increase permeability of cell membrane
4-cell damage with the release of its enzyme content
1- bilirubin increases the rate of synthesis of alkaline phosphatase in obstructive liver diseases
2- obstruction of bile ducts increases alkaline phosphatase
3-as in tissue hypoxia
4-cell damage with the release of its enzyme content into the blood like myocardial infarction and viral hepatitis
medical importance of measuring the non functional plasma enzymes 2
diagnosis of diseases (detection of diseased organ)
prognosis and follow up the disease before and after treatment
سلايد رقم 37 دا مش عارف اعمل فيه اي انا مش هكتب الاختصارات دي ل ايه بس هي منطقيه يعني
لوجالبلك الكلمه كامله هتفهم انها الاختصار ان شاء الله لو مش هتمشي معاك ذاكر النقطه دي من البي دي اف بقا
examples of medically important non functional plasma enzymes 8
amylase & lipase
CK&LDH
ALT&AST
alkaline phosphatase & acid phosphatase
amylase and lipase increase in
pancreatic diseases
CK increases in
hheart brain and skeletal muscle diseases
LDH increases in
liver blood and heart diseases
ALT increases in
liver diseases
AST increases in
liver heart muscles diseases
acid phosphatase increase in ?
cancer prostate
alkaline phosphatase increase in ?
increase in obstructive liver disease bone disease and hyperparathyroidism
iodine reaction with starch gives color ?
deep blue color
iodine reaction with maltose gives color ?
no color produce
اوعي تتغفل ;)
enzymes activity in normal room temp and if cooled to temp like 2 C or heated until like 100 C
in normal temp works fine
if heated or cooled enzyme wont work
خلصنا اهو