Download - Presentasi Bahan Hayati Laut (BHL)
UJI SITOTOKSIK METODE
MTT (MICROCULTURE TETRAZOLIUM)
BAHAN HAYATI LAUT
KELOMPOK : 5
ROHIMAT
SITI NURUL AINI
YUDHAWIRA B. SEMBIRING
M. NOFRI
1. PENGANTAR
2. METODE MTT ASSAY
3. PENUTUP
?MTT ASSAY
“Salah satu metode
TES SITOTOKSIK yang digunakan untuk menguji
VIABILITAS SEL”
(3-(4,5-dimethiylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)
METHYLTHIAZOL TETRAZOLIUM (MTT)
• Mosmann (1983)
• Berwarna kuning
• Larut dalam airGambar: Struktur MTT
PRINSIP DASAR MTT ASSAY
ENZIME SUCCINE DEHYDOGENASE MITOKONDRIA
METHYLTHIAZOL TETRAZOLIUM (MTT)
Metabolisme sel hidup
KRISTAL FORMAZAN UNGUJumlah sebanding sel hidup
PRINSIP DASAR MTT ASSAY
PRINSIP DASAR MTT ASSAY
• IMPERMEABEL
• TIDAK LARUT DALAM AIR
DMSO + HCL
METODE MTT ASSAY
Gambar. The Epoch™ Multi-Volume Spectrophotometer SystemSumber: BioTek
Inokulasi 96-well plate
1x104 cell/mLDalam Dulbecco’s Modified Eagle’s Medium
(DMEM)
INKUBASI 24 JAM
GANTI MEDIA
METODE MTT ASSAY
Dimethyl Sulfoxide
(DMSO)
0, 1, 12.5, 25, 37.5, 50, 62.5, 75 ng/mL
INKUBASI 3.5 JAM
Phosphate Buffer Saline
(PBS) MTT+
METODE MTT ASSAY
DMSO 150 Mikro L
+
METODE MTT ASSAY
INKUBASI 15 MENIT
FORMAZEN UNGU
Tecan Microplate
reader 590 nm
Filter 620 nm
METODE MTT ASSAY
Sumber: vmsr.net
METODE MTT ASSAY
< 100% = Toksik
“The MTT assay provides a safe, effective method to quantify cell viability or proliferation in response to a variety
of external stimuli”Peter Brescia, M.Sc. and Peter Banks, Ph.D., BioTek Instruments,
Inc
PENUTUP
TERIMA KASIH