Measuring Protein Expression and Cellular Fluorescence [Mean Gray Value vs Integrated Density]

Описание: Learn how to use FIJI (ImageJ) to correctly measure cellular fluorescence and protein expression, and make the choice on whether to report mean fluorescence or total fluorescence based on the biological question you want to answer. Taking out background measurements as well as normalizing measurements also need to be accounted for. The examples shown in this tutorial are for 2D images.

Timeline:
00:00 Introduction
00:06 A. Measuring the amount of protein expressed inside a cell/object
03:29 B. Measuring cell fluorescence: Corrected Total Cell Fluorescence
07:02 C. Measuring protein concentration

Image credits:
A. Tiantian Lin, Cornell University (lab of Alireza Abbaspouurad, Dept. of Food Science)
B. Ashley Cardenas, Cornell University (lab of Lawrence Bonassar, Meinig School of Biomedical Engineering)
C. Hung-Hsiang Huang, Pamela Stanley (2011): CIL13656 and CIL13657
(Cell Image Library: https://doi.org/doi:10.7295/W9CIL13656, https://doi.org/doi:10.7295/W9CIL13657)

LUTS used in images:
A. KTZ_bt_Teal and KTZ_bt_Brick (KTZ-LUTs update site in Fiji, from Kevin Terretaz)
B. JDM Duo Yellow (James Manton LUTs)
C. k_Green and k_Orange

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