Micro incubator

Manufactured by Okolab

Sourced in Italy

The Micro-incubator is a compact and precise temperature control system designed for microscopy applications. It maintains a stable and uniform temperature environment within a small enclosed chamber, ensuring optimal conditions for live-cell imaging and analysis.

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Lab products found in correlation

Symphotime 64, by PicoQuant (2 mentions) Ldh d c 485, by PicoQuant (1 mentions) P35g 0.170 14 c, by MatTek (1 mentions) A1 scanning confocal microscope, by Nikon (1 mentions) Dic plan apochromat vc objective, by Nikon (1 mentions) Fluorobrite medium, by Thermo Fisher Scientific (1 mentions) Time correlated single photon counting module, by PicoQuant (1 mentions) Eclipse ti a1r microscope, by PicoQuant (1 mentions) 4.2 plus camera, by Oxford Instruments (1 mentions) Csu w1, by Yokogawa (1 mentions)

3 protocols using micro incubator

Lysosome Dynamics Imaging in Cells

Cited 1 time

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The cells were grown on 35 mm glass bottom dishes (Mattek Corporation, P35G-0.170 14-C). 24 h after seeding, cells were incubated for 30min with 1μM Lyso Flipper^{53 (link)} in fluorobrite medium (Gibco, 21083027) supplemented with 10% FCS (without FCS for HPDE) for 30min before imaging. For live imaging, cells were maintained at 37°C and 5% CO2 in a micro-incubator (Okolab, Pozzuoli NA, Italy). LLOMe was used at 1mM. For FLIM imaging, a 100× 1.45 NA oil DIC Plan-Apochromat VC objective (Nikon) with a Nikon A1 scanning confocal microscope was used. Excitation was performed using a pulsed laser at 485 nm (PicoQuant, LDH-D-C-485) at 20 MHz, and the emission signal was collected between 550 and 650 nm using a gated PMA hybrid detector and a TimeHarp 260 Nano Dual TSCPC unit. FLIM images were analyzed using the magic wand tool in SymPhoTime 64 software (PicoQuant) and corresponding pixels were fitted with a dual exponential reconvolution model whereby the lifetime τ1 was extracted. Data are expressed as means ± SD.

Gupta S., Yano J., Mercier V., Htwe H.H., Shin H.R., Rademaker G., Cakir Z., Ituarte T., Wen K.W., Kim G.E., Zoncu R., Roux A., Dawson D.W, & Perera R.M. (2021). Lysosomal retargeting of Myoferlin mitigates membrane stress to enable pancreatic cancer growth. Nature cell biology, 23(3), 232-242.

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Membrane Tension Evaluation by FLIM

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Membrane tension was evaluated by FLIM of polarized CFTR-CTL and CFTR-KD cultures incubated for 1 h at the basal side with the FliptR fluorescent probe (1 μl/ml, SC020; Spirochrome) under controlled temperature and atmosphere (37°C, 5% CO₂). FLIM imaging was performed using a Nikon Eclipse Ti A1R microscope equipped with a Time-Correlated Single-Photon Counting module from PicoQuant, as previously described (Colom et al, 2018 (link)), and a water immersion Apo LWD 40X/1.15 N.A. objective (Nikon). During acquisitions, cells were maintained at 37°C and 5% CO₂ with a micro-incubator (Okolab). FliptR fluorescence lifetime was determined for at least 10 positions, always at the same height within each Transwell filter. SymPhoTime 64 software (PicoQuant) was used to fit fluorescence decay data (from full images or regions of interest) to a dual exponential model after deconvolution for the instrument response function (measured using the backscattered emission light of a 1 μM fluorescein solution with 4M KI). Data were expressed as the mean ± SD of the mean.

Simonin J.L., Tomba C., Mercier V., Bacchetta M., Idris T., Badaoui M., Roux A, & Chanson M. (2024). Apical dehydration impairs the cystic fibrosis airway epithelium barrier via a β1-integrin/YAP1 pathway. Life Science Alliance, 7(4), e202302449.

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Time-lapse Imaging of Actin-labeled Cells

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Time-lapse imaging was performed with an inverted microscope Nikon Ti-E installed into a thermostatically controlled chamber (Life Imaging Technologies) and equipped with a micro-incubator for thermal, CO₂ and humidity control (OKOlab). The microscope was also equipped with an automated stage and a Yokogawa CSU-W1 spinning disk unit. Image acquisition was performed with an Andor Zyla 4.2 Plus camera, operated with Slidebook (ver.6.0.19). We performed fluorescence (60x lens, NA 1.4), phase contrast (10/20x objectives, NA 0.3/0.45) and differential interference contrast (DIC) imaging (20x lens, NA 0.45). 4D time-lapse was used for actin-labelled cell mounds (60x lens, NA 1.4) and for the pillar compression experiments. The latter combined DIC and confocal fluorescence modes (20x lens, NA 0.45). Typically, we acquired 12 images/h for >10h.

Guillamat P., Blanch-Mercader C., Pernollet G., Kruse K, & Roux A. (2022). Integer topological defects organize stresses driving tissue morphogenesis. Nature materials, 21(5), 588-597.

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