CellPaint™ TSP membrane stain

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	1016.27
Solvent	DMSO

Spectral properties

Excitation (nm)	496
Emission (nm)	633

Storage, safety and handling

Intended use	Research Use Only (RUO)
Storage	Freeze (< -15 °C); Minimize light exposure

Overview SDS Protocol

Molecular weight

1016.27

Excitation (nm)

496

Emission (nm)

633

TSP is a styrylpyridine-based fluorescent membrane probe suitable for imaging plasma membranes in living cells and tissues. It was reported by Guo et al in 2016 (Analyst, 2016, 141, 3228). The probe is a molecular rotor that has fluorescence sharply enhanced in viscous media. Its fluorescence is also microenvironment-sensitive, enables the turn-on imaging of plasma membranes with a high signal-to-noise ratio. Guo et al has demonstrated that TSP has high photostability, low cytotoxicity and excellent biocompatibility. It can also be used in 2 photon imaging.

Platform

Fluorescence microscope

Excitation	Cy3/TRTC filter set
Emission	Cy3/TRITC filter set
Recommended plate	Black wall/clear bottom

Example protocol

AT A GLANCE

Prepare cells in growth medium
Incubate cells with Cellpaint™ TSP membrane stain
Analyze under fluorescence microscope with Cy3/TRITC filter set

Chemical and Physical Properties

Molecular Weight: 1016.27
Solvent: dimethyl sulfoxide (DMSO)
Spectral Properties: Excitation = 490 nm; Emission = 600 nm

CELL PREPARATION

For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

Cellpaint™ TSP membrane stain stock solution (500X)

Thaw the vial of CellPaint™ TSP membrane stain at room temperature before use.
Add 100 µL of DMSO into the vial of Cellpaint™ TSP membrane stain to make 500X stock solution.
Note Unused Cellpaint™ TSP membrane stain stock solution can be aliquoted and stored at ≤ -20 °C for a couple of months if the tubes are sealed tightly. Protect from light and avoid repeated freeze-thaw cycles.

PREPARATION OF WORKING SOLUTION

Cellpaint™ TSP membrane stain working solution (10X)

Add 20 µL of 500X stock solution into 1 mL of cell culture medium, and mix well.
Note     We recommend making the working solution fresh before use.
Note     20 µL of 500X Cellpaint™ TSP membrane stain stock solution is enough for one 96-well plate.
Note     We recommend using a cell culture medium to make a working solution for better resolution images.

SAMPLE EXPERIMENTAL PROTOCOL

Stain Cells

Add 10 µL/well (96-well plate/100 µL volume) or 5 µL/well (384-well plate-50 µL volume) of 10X Cellpaint™ TSP membrane stain working solution in the cell plate. Incubate the cells at 37 °C for 30-60 minutes, protected from light.
Note The optimal concentration of the cell membrane probe varies depending on the specific application. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe.
Remove working solution in each well. Wash cells with physiological buffer (such as HHBS, DPBS or buffer of your choice) for three times and replace with HHBS.
Observe the fluorescence signal in cells using fluorescence microscope with a Cy3/TRITC filter set.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of CellPaint™ TSP membrane stain to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	98.399 µL	491.995 µL	983.99 µL	4.92 mL	9.84 mL
5 mM	19.68 µL	98.399 µL	196.798 µL	983.99 µL	1.968 mL
10 mM	9.84 µL	49.2 µL	98.399 µL	491.995 µL	983.99 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	496
Emission (nm)	633

Images

Figure 1. HeLa cells were labeled with CellPaint™ TSP membrane stain (Cat No. 22700). Labeled cells were imaged on the Keyence BZ-X710 All-In-One Fluorescence Microscope equipped with a Cy3/TRITC filter set.

Figure 2. Chemical structure for CellPaint™ TSP membrane stain.

Citations

View all 31 citations: Citation Explorer

Membrane trafficking and exocytosis are upregulated in port wine stain blood vessels
Authors: Yin, R., Rice, S. J., Wang, J., Gao, L., Tsai, J., Anvari, R. T., Zhou, F., Liu, X., Wang, G., Tang, Y., Mihm, M. C., Jr., Belani, C. P., Chen, D. B., Nelson, J. S., Tan, W.
Journal: Histol Histopathol (2019): 479-490

Cu(2+)-Directed Liposome Membrane Fusion, Positive-Stain Electron Microscopy, and Oxidation
Authors: Liu, Y., Liu, J.
Journal: Langmuir (2018): 7545-7553

Extraction of DNA from Sperm Cells in Mixed Stain by Nylon Membrane Bushing Separation Technique
Authors: Ma, J., Tong, Q., Gao, L. B., Zhu, C., Jiang, Z. Q.
Journal: Fa Yi Xue Za Zhi (2018): 417-419

Gram's Stain Does Not Cross the Bacterial Cytoplasmic Membrane
Authors: Wilhelm, M. J., Sheffield, J. B., Sharifian Gh, M., Wu, Y., Spahr, C., Gonella, G., Xu, B., Dai, H. L.
Journal: ACS Chem Biol (2015): 1711-7

The use of SMALPs as a novel membrane protein scaffold for structure study by negative stain electron microscopy
Authors: Postis, V., Rawson, S., Mitchell, J. K., Lee, S. C., Parslow, R. A., Dafforn, T. R., Baldwin, S. A., Muench, S. P.
Journal: Biochim Biophys Acta (2015): 496-501

Overview of electron crystallography of membrane proteins: crystallization and screening strategies using negative stain electron microscopy
Authors: Nannenga, B. L., Iadanza, M. G., Vollmar, B. S., Gonen, T.
Journal: Curr Protoc Protein Sci (2013): Unit17 15

Negative-stain electron microscopy of inside-out FtsZ rings reconstituted on artificial membrane tubules show ribbons of protofilaments
Authors: Milam, S. L., Osawa, M., Erickson, H. P.
Journal: Biophys J (2012): 59-68

Ultrastructure and retinal imaging of internal limiting membrane: a clinicopathologic correlation of trypan blue stain in macular hole surgery
Authors: Mackenzie, S. E., G and orfer, A., Rohleder, M., Schumann, R., Schlottmann, P. G., Bunce, C., Xing, W., Gregor, Z., Charteris, D. G.
Journal: Retina (2010): 655-61

A nitrobenzofuran-conjugated phosphatidylcholine (C12-NBD-PC) as a stain for membrane lamellae for both microscopic imaging and spectrofluorimetry
Authors: Hope-Roberts, M., Wainwright, M., Horobin, R. W.
Journal: Biotech Histochem (2008): 25-8

Indocyanine green-assisted internal limiting membrane peeling for macular holes to stain or not to stain?
Authors: Da Mata, A. P., Riemann, C. D., Nehemy, M. B., Foster, R. E., Petersen, M. R., Burk, S. E.
Journal: Retina (2005): 395-404