An Introduction to Performing Immunofluorescence Staining

Abstract

Summary Immunofluorescence (IF) is an important immunochemical technique that allows detection and localization of a wide variety of antigens in different types of tissues of various cell preparations. IF allows for excellent sensitivity and amplification of signal in comparison to immunohistochemistry, employing various microscopy techniques. There are two methods available, depending on the scope of the experiment or the specific antibodies in use: Direct (Primary) or Indirect (Secondary). Here, we describe preparation of specimens preserved in different types of media and step-by-step methods for both direct and indirect immunofluorescence staining.

The CoViBE: An Innovating Self-Paced Elearning to Teach Virtually Bench-Top Practice

“…Immunocytofluorescence is a basic technique in cell biology consisting of "staining" a cellular protein using specific antibodies (Im et al, 2019). In our case, highlighted proteins were vimentin (expressed in cell cytoskeleton) and nuclear laminin (expressed in nucleus).…”

“…Actually, emerging research shows that comics and graphic novels are motivating, support struggling readers, enrich the skills of accomplished readers and are highly effective at teaching sometimes dull or dry material in subject areas such as science and social studies (Hosler & Boomer, 2011;MANNO, 2014;McCroskey & Richmond, 1983;Smetana et al, 2009). This first generation of CoViBE is about immunocytofluorescence (Im et al, 2019), a highly used technique in biology and so a usual bench-top technique concerning students at the undergraduate program of Bordeaux University (second year of Bachelor"s degree in life sciences). The aim of this paper deals with the use of comics for teaching practical work sessions by distance and the study of its influence on student"s learning.…”

P2X7 blockade inhibits the growth of breast cancer in 4T1 breast cancer-bearing mice by NLRP3/caspase 1 pathway

“…The immunofluorescence staining was conducted in concordance with the related guidelines [26]. Generally, the tumour sections were incubated with primary antibodies overnight at 4°C and were further covered with secondary antibodies (Jackson Immunoresearch, West Grove, PA) for 2 h at room temperature accordingly.…”

A 3D mixing-based portable magnetic device for fully automatic immunofluorescence staining of γ-H2AX in UVC-irradiated CD4+ cells

“…The light-tight plastic housing and support components were three-dimensionally printed in the laboratory. The assembled magnetic device features a size of 192 Â 123 Â 122 mm 3 (Fig. 3), a weight about 0.666 kg and a power consumption less than 10 W, with a built-in Single Chip Microcomputer (SCM, STC12C2052AD) for control of three stepper motors (one rotary motor and two moving motors) to fully automate the mixing and movement of MBs ( Fig.…”

“…Immunouorescence (IF) is a highly sensitive and specic method that has been widely used in cell biology to study differential expression, localization and distribution of proteins at the tissue, cellular, and subcellular level. [1][2][3] It has been regarded as a routine technique in basic biomedical research and an effective approach for biomarker diagnostics. [4][5][6] The nucleosomal histone protein H2AX is specically phosphorylated (g-H2AX) adjacent to DNA double-strand breaks (DSBs) and is used for quantifying DSBs.…”