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