Cell Staining　DOJINDO LABORATORIES

Classification

Cell Staining

Introduction

Visualization of a cell with fluorescent compounds provides a wide variety of information for the analysis of cell functions. Various activities and structures of a cell can be targeted for staining with fluorescent compounds (Fig. 1). The most commonly stained cell components are cell membranes, proteins, and nucleotides. Small neutral molecules and positively charged molecules can pass through viable cell membranes and remain inside of cells, depending on their reactivity or hydrophilicity. Negatively charged molecules cannot pass through viable cell membranes. Positively charged molecules are usually cell membrane permeable and accumulate in mitochondria. Ester is a suitable functional group for staining viable cells. Ester can pass through viable cell membranes, where it is hydrolyzed by cellular esterases into a negatively charged molecule under physiological conditions. Several fluorescein analogs with ester groups in their structure are available for staining viable cells. Succinimidyl ester compounds can also be used to improve retention of the fluorescent derivative within the cell. These compounds are neutral molecules that pass through cell membranes and covalently conjugate with cell proteins. Covalently conjugated molecules can stay in the cell for several weeks. Nucleotide staining with fluorescent intercalators is mostly used for dead cell detection.

Cell Cytosol Staining

Fluorogenic esterase substrates that can be passively loaded into viable cells, such as Calcein-AM, BCECFAM, Carboxyfluorescein succinimidyl ester (CFSE), and Fluorescein diacetate (FDA), are converted by intracellular esterases into fluorescein analogs with green fluorescence. Calcein and BCECF can be converted into electrically neutral molecules by the addition of acetyl or acetoxymethyl groups to their phenolic OH or carboxylic groups, which allows them to freely permeate into the cell. Once converted into fluorescent products by esterase, these compounds are retained by cells because of their negative charges. These esterase substrates, therefore, can serve as assay probes of cell viability. Fluorescent esterase substrates may also be used in cell viability assays in place of tetrazolium analogs such as MTT or WST. The mechanism for determining cell viability is different: Although both assays determine cell metabolism, esterase substrates detect esterase activity; tetrazolium salts detect dehydrogenase activity of viable cells. CFSE is also an ester compound that passes through viable cell membranes. Since it has an amine-reactive succinimidyl group, fluorescein derived from CFSE can covalently bond to proteins or other amino groups in the cell or on the cell membrane. This covalently attached fluorescein remains stable and allows the cell to be traced over several weeks.

Mitochondria Staining

Mitochondria exist in most eukaryotic cells and play a very important role in oxidative metabolism by generating ATP as an energy source. The average number of mitochondria per cell is from 100 to 2,000. Although the typical size is about 0.5-2 mm, the shape, abundance, and location of mitochondria vary by cell type, cell cycle, and cell viability.Therefore, visualization of mitochondra is important. Since mitochondria have electron transport systems, they can be stained with various redox dyes. MitoRed and Rh123 readily pass through cell membranes and accumulate in mitochondria. The fluorescence intensity of Rh123 reflects the amount of ATP generated in mitochondria.

Nucleus Staining

Fluorescent dyes with aromatic amino or guanidine groups, such as propidium iodide (PI), diaminophenylindole (DAPI), acridine orange (AO), and Hoechst dyes, interact with nucleotides to emit fluorescence. PI molecules intercalate inside the DNA double helix. DAPI and Hoechst dye molecules attach at the minor groove of the DNA double helix. On the other hand, AO can form complexes with either double-stranded DNA or single-stranded DNA and RNA. One molecule of AO can intercalate with three base pairs of double-stranded DNA to emit green fluorescence with the maximum wavelength at 526 nm. One molecule of AO can also interact with one phosphate group of single-stranded DNA or RNA to form an aggregated, or stacked, structure that emits red fluorescence with the maximum wavelength at 650 nm. Cell membranes of viable cells are impermeable to these fluorescent dyes,except for the Hoechst dyes, and these dyes can therefore be used as fluorescent indicators of dead cells. Hoechst dyes are positively charged under physiological conditions and can pass through viable cell membranes.

Bacterial Cell Staining

There are several ways to detect bacteria, including agar plate cultivation and bacteria-specific DNA amplification. Fluorescent staining using CTC is another method used to detect viable bacterial cells. The advantage of this method is very quick detection and the possibility of detecting VNC (viable but non-culturable) bacterial cells. CTC is a tetrazolium salt that is converted to formazan dye by bacterial cell activity. The solid state of the formazan dye emits red fluorescence. Therefore, viable bacterial cells can be stained by CTC and are easily detected by fluorescent microscopy.

Product

Cell Double Staning Kit

Code	Product name	Unit size
CS01	Cell Double Staining -Cellstain- Double Staining Kit	1 set

Live Cell Staining

Code	Product name	Unit size
C326	Live Cell Staining -Cellstain- Calcein-AM	1 mg
C375	Live Cell Staining -Cellstain- CFSE	1 mg
C396	Live Cell Staining -Cellstain- Calcein-AM solution	1 ml
C410	Live Cell Staining -Cellstain- CytoRed solution	1 ml
F209	Live Cell Staining -Cellstain- FDA	1 mg

Dead Cell Staining

Code	Product name	Unit size
C555	Dead Cell Staining (Blue) Dead Cell Makeup Blue - Higher Retention than PI	100 tests
D212	Dead Cell Staining -Cellstain- DAPI	1 mg
D523	Dead Cell Staining -Cellstain- DAPI solution	1 ml
P346	Dead Cell Staining -Cellstain- PI	1 mg
P378	Dead Cell Staining -Cellstain- PI solution	1 ml

Nuclear Staining

Code	Product name	Unit size
A430	Nucleus Dye -Cellstain- AO solution	1 ml
H342	Nucleus Dye -Cellstain- Hoechst 33342 solution	1 ml

Mitochondria Staning

Code	Product name	Unit size
M466	Mitochondrial Lipid Peroxide Detection MitoPeDPP	5 ug x 3
M489	Mitochondrial Iron Detection Mito-FerroGreen	1 set(50 ug x 2)
MD01	Mitophagy Detection Mitophagy Detection Kit	1 set
MT02	Mitophagy Detection Mtphagy Dye	5 μg x 3
MT05	Mitochondrial Singlet Oxygen Imaging Si-DMA for Mitochondrial Singlet Oxygen Imaging	2 ug
MT09	Mitochondrial Membrane Potential Detection JC-1 MitoMP Detection Kit	1 set
MT10	Mitochondrial Staining MitoBright LT Green	400 ul
MT11	Mitochondrial Staining MitoBright LT Red	400 ul
MT12	Mitochondrial Staining MitoBright LT Deep Red	400 ul
MT13	Mitochondrial Membrane Potential Detection MT-1 MitoMP Detection Kit	1 set
MT15	Mitochondria Fluorescent Probe for Immunostaining Red MitoBright IM Red for Immunostaining	20 ul 20 ul x 3
MT16	Mitochondrial Superoxide Detection MitoBright ROS Deep Red - Mitochondrial Superoxide Detection	100 nmol x 1 100 nmol x 3
R237	Mitochondrial Staining -Cellstain- MitoRed	50 ug x 8

Tissue Staining

Code	Product name	Unit size
D006	Coloring Substrate for Peroxidase DAB	100 mg 1 g 5 g
F308	Amyloid Staining FSB solution	100 ul
T022	Coloring Substrate for Peroxidase TMBZ	1 g 5 g
T039	Coloring Substrate for Peroxidase TMBZ･HCl	1 g

Nucleolus Staining

Code	Product name	Unit size
N511	Nucleolus Fluorescent Staining Nucleolus Bright Green	60 nmol
N512	Nucleolus Fluorescent Staining Nucleolus Bright Red	60 nmol

Lipid Droplet Staining

Code	Product name	Unit size
LD01	Lipid Droplet Staining Lipi-Blue	10 nmol x 1
LD02	Lipid Droplet Staining Lipi-Green	10 nmol x 1
LD03	Lipid Droplet Staining Lipi-Red	100 nmol x 1
LD04	Lipid Droplet Staining Lipi-Deep Red	10 nmol *
LD05	Lipid Droplet Assay Lipid Droplet Assay Kit - Blue	1 set *
LD06	Lipid Droplet Assay Lipid Droplet Assay Kit - Deep Red	1 set *

Cell Membrane Staining

Code	Product name	Unit size
E296	Endocytosis Detection ECGreen-Endocytosis Detection	40 μl
P504	Cell Membrane Staining PlasMem Bright Green	100 ul
P505	Cell Membrane Staining PlasMem Bright Red	100 ul

Lysosome Staining

Code	Product name	Unit size
L261	Lysosome Staining Dye Green LysoPrime Green - High Specificity and pH Resistance	10 ul x 1 10 ul x 3
L264	Lysosome Staining Deep Red LysoPrime Deep Red - High Specificity and pH Resistance	1 tube 3 tubes
L265	Lysosomal pH Detection Reagent Red pHLys Red - Lysosomal Acidic pH Detection	1 tube 3 tubes
L266	Lysosomal Acidic pH Detection Kit - Green/Red Lysosomal Acidic pH Detection Kit	1 set
L268	Lysosomal Acidic pH Detection Kit-Green/Deep Red Lysosomal Acidic pH Detection Kit-Green/Deep Red	1 set

Classification

Cell Staining

Introduction

Cell Cytosol Staining

Mitochondria Staining

Nucleus Staining

Bacterial Cell Staining

Product

Product Classification

Product Classification

Cell Proliferation / Cell Cytotoxicity Assay

Cell Staining

Intracellular Fluorescent Probes

Labeling Chemistry

Oxidative Stress

-Bacstain- Series

Molecular Biology

Detergents

Cross-Linking Reagents

Redox Dyes

Ion Analysis

Buffers

Metal Chelates

Chromogen/Metal Indicator

Biochemicals

Functional Organic Material