Lsm 510 meta microscope

Manufactured by Nikon

The Nikon LSM 510 Meta is a high-performance laser scanning confocal microscope. It features a multi-track imaging system and can detect multiple fluorescent labels simultaneously. The microscope is equipped with a range of laser lines and detection channels to enable advanced imaging capabilities.

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Lab products found in correlation

A1 laser scanning microscope, by Nikon (2 mentions)

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LSM 510 (16 citations) LSM 510 META (6 citations)

3 protocols using lsm 510 meta microscope

Cas9En-RNP:ArgNPs Nanoassembly Delivery

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Assembled Cas9En-RNP:ArgNPs nanoassemblies (preassembled in 100 μL PBS for 10 min, plus 900 μL DMEM media) were immediately transferred to each dish of confluently grown cells. Cells were then incubated at 37 °C and 5% CO₂ for 3h. At this point cells were washed with 1xPBS buffer and immediately processed for investigating delivery efficiency that was determined by Confocal microscopy (Zeiss LSM 510 Meta microscope, or Nikon A1 laser scanning microscope). Z-stacking was performed using Nikon A1, at every 125 nm interval.
It is noteworthy that the nanoassemblies can be incubated with cells for a longer period of time (~24h) without affecting cell growth/viability, an important issue for in vitro and ex vivo editing.

Mout R., Ray M., Tonga G.Y., Lee Y.W., Tay T., Sasaki K, & Rotello V.M. (2017). Efficient Gene Editing through Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein. ACS nano, 11(3), 2452-2458.

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Confocal Microscopy for Cellular Uptake

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Confocal microscopy (Zeiss LSM 510 Meta microscope or Nikon A1 laser scanning microscope) was utilized for evaluating the cellular uptake of proteins. After each delivery process, the cells were washed with PBS three times to remove free proteins. Then cells were then fixed with a 4% paraformaldehyde aqueous solution for 20 min at room temperature and rinsed with PBS. For co-localization of delivered payload, cells were stained with DAPI at room temperature for 10 min and rinsed with PBS. Afterwards, delivery efficiency was determined by confocal microscopy. Z-stacking was conducted using Nikon A1, at every 125 nm interval. The delivery efficiency was estimated by mean fluorescence intensity (MFI) of confocal images (as shown in Figure S3), which were further analyzed by NIS-Elements software and plotted in histogram form.

Pan S., Jeon T., Luther D.C., Duan X, & Rotello V.M. (2020). Cytosolic Delivery of Functional Proteins In Vitro through Tunable Gigahertz Acoustics. ACS applied materials & interfaces, 12(13), 15823-15829.

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Multispecies Biofilm Imaging Assay

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GFP-expressing MRSA and DsRed-expressing E. coli were inoculated in Luria broth (LB) medium at 37 °C until the stationary phase. Cultures were harvested by centrifugation and washed with 0.85% sodium chloride solution three times. Concentrations of resuspended bacterial solutions were determined by optical density measured at 600 nm. A quantity of 10⁸ bacterial cells /mL of GFP-expressed MRSA, DsRed-expressing E. coli or their combination, supplemented with 1 mM of isopropyl β-d-1-thiogalactopyranoside (IPTG), were seeded (2 mL in TSB) in a confocal dish and were allowed to grow. After 4 days, media were replaced by M9 media containing 1 μM of coumarin blue-tagged PNPs, then incubated for 1 h. Biofilm samples incubated with M9 media only were used as controls. Cells were imaged before and after washing with PBS three times. Confocal microscopy images were obtained on a Zeiss LSM 510 Meta microscope, from the Light Microscopy Facility and Nikon Center of Excellence at the Institute for Applied Life Sciences, UMass Amherst, by using a 40× objective. The settings of the confocal microscope were as follows: green channel, λ_ex = 488 nm and λ_em = LP 540 nm; red channel, λ_ex = 560 nm and λ_em = LP 640 nm; blue channel, λ_ex = 403 nm and λ_em = LP 495 nm. Emission filter: LP = high pass.

Makabenta J.M., Park J., Li C.H., Chattopadhyay A.N., Nabawy A., Landis R.F., Gupta A., Schmidt-Malan S., Patel R, & Rotello V.M. (2021). Polymeric Nanoparticles Active against Dual-Species Bacterial Biofilms. Molecules, 26(16), 4958.

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