PKmem 590 is a bright, non-phototoxic & non-toxic plasma membrane probe based on the PKmem™ dyes developed by the lab of Zhixing Chen at Peking University1. PKmem 590 labels the plasma membrane in live cells with very high specificity. The unique and unmatched feature of PKmem 590 is its extremely low phototoxicity, due to the presence of the intramolecular triplet quencher cyclooctatetraene (COT) group. It allows to perform long term imaging the plasma membrane without damaging the cells. It is highly suited to image plasma membrane structure and dynamics by STED superresolution microscopy using a 775 nm depletion line. PKmem 590 does not require any genetic manipulation, transfection or overexpression of fluorescent proteins. PKmem 590 enables multicolor imaging with SPY505, SPY555, SPY620, SPY650, SPY700, SiR or GFP. It can be used for widefield, confocal, SIM or STED imaging in living cells and tissue. Contains 1 vial of PKmem 590 (lyophilized).
Absorbance Maximum λabs (MeOH) | 590 nm |
Fluorescence maximum λfl (MeOH) | 606 nm |
Works on fixed cells? | yes |
Probe quantity | 100 stainings* |
Fluorescence lifetime | n.d. |
STED depletion wavelength | 775 nm |
Shipping | room temperature |
Storage | -20°C |
mESC_PKmem590_PKmitoDEEPRED
PKmem590_HeLa
For product Datasheets and MSDSs please click on the PDF links below.
Spirochrome Technical Tips and Ex/Em spectra in graphical form (PDF)
If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com
Q1. What is STED microscopy and how does it work?
A1. STED microscopy stands for Stimulated Emission Depletion microscopy. It is one type of super resolution microscopy which allows the capture of images with a higher resolution than conventional light microscopy which is constrained by diffraction of light. STED uses 2 laser pulses, one is the excitation pulse which excites the fluorophore, causing it to fluoresce. The second pulse, referred to as the STED pulse, de-excites the fluorophore via stimulated emission in an area surrounding a central focal spot that is not de-excited and thus continues to fluoresce. This is accomplished by focusing the STED pulse into a ring shape, a so-called donut, where the center focal spot is devoid of the STED laser pulse, conferring high resolution to the fluorescent area (Fig. 1; see Ref. 1 for more details on STED microscopy).
Figure 1. STED microscopic image of microtubules labeled with SiR-tubulin in human primary dermal fibroblasts.
Q2: Are the SPY™ probes stable at room temperature?
A2: Yes, the probes are stable at room temperature for a few days. However, it strongly depends on the probe and the solvent. Thus, it is recommended to store all of the probes or solutions at –20°C.
References
1. Liu Tianyan et al. “Multi-color live-cell STED nanoscopy of mitochondria with a gentle inner membrane stain” PNAS (2022):119 (52).
2. Yang, Zhongtian, et al. “Cyclooctatetraene-conjugated cyanine mitochondrial probes minimize phototoxicity in fluorescence and nanoscopic imaging.” Chemical science 11.32 (2020): 8506-8516.
