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[Fluorescence probe for super-resolution live imaging] HMSiR

  • Available for live cell imaging under the physiological condition
  • Blinking without high-power laser irradiation

Super-resolution imaging techniques, that operate beyond the limit set by the diffraction of light, obtains many new knowledge in cellular biology. Among these techniques, dSTORM (direct stochastic optical reconstruction microscopy), that utilizes blinking fluorescent dyes, are widely known as the method allowing spatial resolution of about 20 nm. The new released HMSiR displays spontaneously blinking in light emission. By using this probe, we can operate super-resolution imaging without thiols, oxygen scavengers or irradiation of high-power laser, all of which were necessary for causing the blinking of fluorescent dyes in the previous dSTORM observation. Therefore, this probe enables live-cell and super-resolution imaging with low-power excitation light under the physiological condition.

Feature of HMSiR

1. Available for live cell imaging under the physiological condition

Fig. 1. Super-resolution and live image of microtubules in Velo cell. Fig. 1. Super-resolution and live image of microtubules in Velo cell.

Velo cells, in which HaloTag®-β-tubulin was expressed, were stained by HMSiR-Halo and observed in the medium of Leibovitz L15 containing 10%FBS after replating. Images were obtained by STORM microscopy (N-STORM:TIRFM:647 nm, 100 W/cm2, 15 ms/frame, 1000 frame). A: averaged image. B: super-resolution image. C: Measurement of localization precision in the same laser condition of Fig. 1A and B. At first, antibodies were labeled with HMSiR, absorbed on the glass surface, and observed by STORM (N-STORM). Positions of the molecule were repeatedly obtained and their distribution was fitted by Gaussian. Only signals over 150 photons were analyze. 647 nm, 100 W/cm2, 15 ms/frame, 10 mM PBS (pH7.4). Super-resolution imaging with HMSiR did not need thiols or oxygen scavengers and was operated under the physiological condition. HaloTag® is a trademark of Promega Corporation.

2. Blinking spontaneously without high-power laser irradiation

Fig. 2. Reaction of HMSiR. Fig. 2. Reaction of HMSiR.

Fig. 3. Fluorescence from single HMSiR molecule.

Fig. 3. Fluorescence from single HMSiR molecule.

HMSiR displays spontaneously blinking without irradiation of high-power laser (~1 kW cm-2) which is used to be necessary for the dSTORM observation (Fig. 3, laser power: 100 W cm-2, correspondence to On/Off state in Fig. 2). It allows us to minimize the damage of the cell by the laser irradiation used in the previous dSTORM observation.

Please refer to the application note.

Reference
Uno SN, Kamiya M, Yoshihara T, Sugawara K, Okabe K, Tarhan MC, Fujita H, Funatsu T, Okada Y, Tobita S, Urano Y. Nat Chem. 2014 681-689.
Line-up of the probes for the super-resolution imaging
HaloTag® is a trademark of Promega Corporation. If you are interested in our services and products, please contact us, E-mail: info_itnl@goryochemical.com.
Applications Code Products Size Price Remarks
For live cells A201-01 HMSiR-Halo 15 nmol $897 Affinity ligands to HaloTag® fusion proteins ( In preparation )

A201-02 30 nmol $1,497
For fixed cells A202-01 HMSiR labeled Goat anti-mouse IgG (whole) 100 μg $897

IgG antibody, from goat, labeled by HMSiR

For fixed cells A203-01 HMSiR labeled Goat anti-rat IgG (whole) 100 μg $897
For fixed cells A204-01 HMSiR labeled Goat anti-rabbit IgG (whole) 100 μg $897
Amine-reactive probe A208-01 HMSiR-NHS 100 μg $747 Labeling amines in proteins and other compounds.

Thiol-reactive probe A209-01 HMSiR-Maleimide 100 μg $747 Labeling thiols in proteins and other compounds.

Contact Information

For further information or questions on our products, please contact us, E-mail: info_itnl@goryochemical.com