CD 4 FLOWCYTOMETRY

CD4 (cluster of differentiation 4) is

a glycoprotein as cell surface receptors/markers,
found on the surface of immune cells such as T
helper cells, monocytes, macrophages,
and dendritic cells.

CD4 is a member of the immunoglobulin

superfamily
• CD4+ T helper cells are white blood cells that
are an essential part of the human immune
system. They are often referred to as CD4
cells, T-helper cells or T4 cells. They are called
helper cells because one of their main roles is
to send signals to other types of immune cells,
which then destroy the infectious particle.
• CD4 cells or T limfosit with CD4 are the main
target of HIV virus
• HIV infection leads to a progressive reduction in the
number of T cells expressing CD4 (CD4 T-cell lysis)
• Medical professionals refer to the CD4 count to decide
when to begin treatment during HIV infection, although
recent medical guidelines have changed to recommend
treatment at all CD4 counts as soon as HIV is diagnosed.
• A CD4 count measures the number of T cells expressing
CD4.
• While CD4 counts are not a direct HIV test—e.g. they do
not check the presence of viral DNA, or specific antibodies
against HIV—they are used to assess the immune system of
a patient.
 CD 4 count
BD FACSCount™ CD4 are
used to enumerate the
absolute counts of CD4 T
lymphocytes and determine
the percentage of
lymphocytes that are CD4 T
lymphocytes in unlysed
whole blood
(CD4 counts and CD4
percentages)
 Prinsip kerja
Darah EDTA + reagen (CD4/CD3 berisi antibodi berlabel florokrom)

Terbentuk ikatan spesifik terhadap sel limfosit

Ditembak dengan sinar laser

Timbul floresen

Pendaran cahaya floresen akan dihitung

 Prinsip kerja
• BDFacscount menggunakan prinsip
flowcytometry  tiap tiap sel pada darah
sampel yang sudah berikatan dengan antibodi
berlabel florokrom akan melewati suatu
tabung  kemudian akan ditembak dengan
sinar laser mengenai label florokrom hingga
berpendar  pendaran cahaya ini akan di
tangkap oleh filter detector 
menggambarkan jumlah sel CD4
 Alat dan bahan
• BP : darah perifer EDTA < 48 jam
• Mesin flowcytometry
• Vortex mixter
• Pipet otomatis
• Tip pipet
• Tabung yang sudah berisi reagen (Antibodi CD4
berlabel florokrom)
• inkubator
• Pembuka tutup tabung
• Larutan fixative
 Langkah - langkah
1. Tabung reagen di putar dahulu pada mesin vortex selama 5-6
detik
2. Lubangi penutup tabung dengan alat pembuka tutup tabung lalu
kesampingkan
3. Masukkan darah sampel sebanyak 50uL lalu di mix dengan mesin
vortex selama 5-6 detik
4. Inkubasi selama 30 menit pada alat inkubator
5. Masukkan larutan fixative sebanyak 50uL lalu di mix lagi dengan
alat vortex selama 5-6 detik
6. Letakkan tabung sampel-reagen pada dudukan tempat sampel
pada alat BDFacscount
7. Alat akan membaca jumlah limfosit T CD4
8. Hasil yang dilaporkan berupa nilai absolut dan persentase