0:01 my name is anish kulkarni i'm a first
0:04 year mbbs student from hbtmc and cooper
0:06 hospital today i'll be performing uh the
0:09 total leukocyte count experiment so
0:11 first we'll check the apparatus for the
0:12 experiment and then we'll i'll tell you
0:20 the basic things we require are spirit
0:22 some cotton
0:24 a lancet
0:27 other than that we require turks fluid
0:30 or the white blood cell diluting fluid
0:33 the nubus chamber
0:40 this is the improved nua so the first
0:42 thing we need to do is sterilize your
0:43 hand to make sure that you do not get
0:45 any infections
0:47 be sure to take the ring finger for for
0:48 any pricking
0:50 procedure as it is not connected to the bursa
1:15 i use the pipette and this uh suck
1:17 suck
1:19 to take the blood inside the pipet up
1:46 if any excess blood enters
1:48 the pivot then use the palm of your hand
1:51 and just tap the pipette till the blood
1:53 is out of the pipette
1:56 then clean off your hand and the tip of
2:08 then add the turks fluid up till the 11
2:24 and then uh use the pipette and suck the
2:26 diluting fluid up till the 11 mark which
2:43 okay now further dial hitting fluid
2:44 until the eleventh point then pull with
2:46 your arms
3:08 then uh
3:10 uh
3:12 take a drop here and then drop it in the
3:14 new pass chamber it will slide across
3:16 the whole numbers chamber due to the
3:31 now let the slide settle for one to two minutes
3:33 minutes
3:34 after you are done charging the new boss
3:36 chamber and you are done waiting for one
3:38 to two minutes uh
3:41 place the nubus chamber in on the microscope
3:43 microscope
3:44 as like this
3:47 then to count use a 10x microscope lens
3:53 to count place this at 10. [Music]
3:55 [Music]
3:57 so this is the diagram for the nubus
4:00 chamber so here here here and here we
4:02 count the white blood cells so start
4:05 from here uh and uh when you see in the
4:07 microscope whatever you see however many
4:09 white blood cells you see start writing
4:10 it here
4:12 and then complete this whole
4:15 chamber dimension so the dimensions of
4:18 the nubus chamber are 1 mm of length and
4:21 bread and 0.1 mm of height so the total
4:23 volume comes to 0.1 mm per so this is
4:26 what the completed nuvar chamber will uh
4:29 diagram will look like with each square
4:30 having some number of white blood cells
4:36 now i'll tell how the dialogue factor of
4:40 20 is achieved so first you take a 0.5
4:43 ml worth of blood and then you add 11 ml
4:45 worth of diluting factor then you
4:47 discard some amount of the
4:49 the
4:51 fluid in the pipette in fluid so if you
4:54 multiply it by 2 then 1 ml of the blood
4:57 contains 20 ml of the diluting fluid
5:04 so now coming to the calculation part
5:06 so the volume of each universe chamber is
5:07 is
5:11 1 into 1 into 0.1
5:14 0.1 mm is the height 1 mm 1 mm is the
5:16 length and 1 mm is the breadth which
5:18 comes down to 0.1 mm cube now this is
5:21 for one box so for four boxes it is
5:23 going to be into four which is equal to
5:26 0.4 mm cube
5:27 of volume now
5:28 now
5:30 after calculating the total number of
5:34 wbcs from these boxes it comes down to 198
5:36 198
5:39 so if 0.4 mm cube
5:43 has 198 wbcs
5:45 so 1 mm cube
5:47 let's take this as x
5:50 so x is going to come out as 198 divided
5:52 by 0.4 now
5:54 now
5:57 this this part won't give us the final
5:59 answer because we'll have to multiply
6:01 this by the diluting factor as well
6:03 which is which is explained before
6:05 and hence after calculating this the
6:08 total number of rbcs will come down as
6:11 9900 per
6:17 so the first question is what is the
6:20 composition of wbc diluting fluid and
6:21 what are the functions of each constrictor
6:23 constrictor
6:25 so the wbc dilating fluid is also known
6:27 as turks fluid it contains glacial
6:28 acetic acid
6:30 which destroys the membranes of our bc
6:32 wbc and platelets the second thing it
6:35 contains its methyl violet which colors
6:37 the fluid and also stains the nuclei of
6:39 the white blood cells and third is
6:40 distilled water which is used as a
6:43 solvent coming to the second question in
6:45 his definition of leukocytosis
6:47 leukopenia and leukemia
6:48 leukemia
6:51 so the leukocytosis is increase in a wbc
6:53 number leukopenia is decreasing wbc
6:56 number and leukemia is excess amount of
6:57 white blood cells in the peripheral
6:59 blood system
7:01 okay the normal range of white blood
7:03 cells is four thousand to eleven
7:05 thousand uh so in leukocytosis uh the
7:08 range uh the number increases above
7:09 eleven thousand that is why it is
7:11 increasing white blood cells number that
7:14 is leukocyte second is leukopenia in it
7:16 uh the white blessing decreases below
7:19 four thousand and third is leukemia in
7:20 which the white blood cell count
7:23 increases well above fifty thousand
7:25 question three is enumerate the causes
7:27 of physiological leukocytosis uh the
7:29 physiological causes are one pregnancy
7:31 or menstruation for women second is
7:32 muscular exercise
7:35 third is diurnal variation and the fourth
7:36 fourth
7:38 is puberty
7:40 pathological causes of leukocytosis
7:43 pathological causes of leukocytes are
7:46 acute and chronic infection the
7:46 the
7:49 allergic reactions and leukemia
7:51 the pyogenic bacterial infection is the
7:53 most common cause of increase in white
7:59 leukocytosis pathological causes of
8:02 leukopenia typhoid fever sulphur drugs
8:04 and irradiation
8:06 and then
8:09 where are the wbcs produced in the body
8:12 the wbcs are produced in the bone marrow
8:15 what is the function of this bead in the
8:17 bulb the first function is it aids in
8:19 mixing of the blood with diluent the
8:21 secondly it helps in identifying the
8:23 pipette as you can see the blade is
8:25 white in color so it is for the white
8:27 blood cells and third it tells whether
8:29 the pipette is dry or not in a dry pet
8:31 the bead will uh
8:33 go around freely but in a wet pipette it
8:35 will stick to the surface and will not
8:37 uh move around freely
8:39 so now we'll discuss the possible
8:41 sources of error and how to minimize
8:43 them one of the sources of error is
8:45 if the pipet is wet
8:47 clean and dry the pipette
8:49 okay and second source of error is if
8:50 blood is
8:53 sucked beyond the 0.5 mark and tap it
8:55 against your palm to go below the 0.5 mark
8:56 mark
8:58 and if there is formation of air bubbles
9:00 while sucking off blood well pick the
9:03 hand carefully and then suck the bell
9:05 and then if if there is sedimentation of
9:08 rbc's in the bulb roll the pipette in
9:09 your palms
9:11 to mix the blood and
9:13 fluid well and why do we need to discard
9:15 the first few drops of rbc because they
9:22 so that's it for the white blood count
