25 × 70 mL puffs of Grape Soda generated at 40 W were vaped onto a #1 coverslip contained in an Attofluor imaging chamber (Thermo-Fisher). Images were then acquired with the 405 nm laser line and collected at 440 ± 20 nm using a Leica SP8 confocal microscope with a 63 × 1.3 NA glycerol immersion lens. A Z-stack of ~60 images were generated and rendered into 3D using Leica LAS Software.
Attofluor imaging chamber
Manufactured by Thermo Fisher Scientific
The Attofluor imaging chamber is a specialized laboratory equipment designed for live-cell imaging applications. It provides a controlled and stable environment for observing and capturing images of live cells under a microscope. The chamber is constructed to maintain optimal temperature, humidity, and gas conditions to support cell viability during imaging sessions.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using attofluor imaging chamber
1
Grape Soda Vapor Imaging
2
Electroformation of Giant Unilamellar Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
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GUVs were made by electroformation.32 (link) Lipid mixtures were dried under vacuum and resuspended
in chloroform, and then a droplet of lipid solution was transferred
to clean indium tin oxide (ITO)-coated slides (Delta Technologies,
Loveland, CO). ITO slides were cleaned by swabbing the slides with
a 2% aqueous Alconox detergent solution and rinsing with Milli-Q H2O immediately thereafter, three times over. Lipid solutions
on the ITO slides were dried under vacuum for a minimum of 15 min
to remove chloroform. A capacitor was formed with a second ITO-coated
slide with a 0.3 mm Teflon spacer. The two slides were sandwiched
together using binder clips. 400 μL of 200 mM sucrose was injected
between the two slides. The total lipid concentration in the chamber
was 1.0 mg/mL. GUVs were electroformed at 65 °C using an AC signal
with a peak-to-peak amplitude of 3.0 V and a frequency of 10 Hz for
2 h (Siglent Technologies). GUVs were removed from the electroformation
chamber and diluted 1:1000 with MOPS buffer in an Attofluor imaging
chamber (Thermo Fisher Scientific). Prior to imaging, GUVs were exposed
to 1 μM EuTc and/or 10 nM CTxB-Alexa647. A minimum of 30 GUVs
among ≥3 individual preparations were examined for signal partitioning,
and representative images were chosen to produce in the figures.
in chloroform, and then a droplet of lipid solution was transferred
to clean indium tin oxide (ITO)-coated slides (Delta Technologies,
Loveland, CO). ITO slides were cleaned by swabbing the slides with
a 2% aqueous Alconox detergent solution and rinsing with Milli-Q H2O immediately thereafter, three times over. Lipid solutions
on the ITO slides were dried under vacuum for a minimum of 15 min
to remove chloroform. A capacitor was formed with a second ITO-coated
slide with a 0.3 mm Teflon spacer. The two slides were sandwiched
together using binder clips. 400 μL of 200 mM sucrose was injected
between the two slides. The total lipid concentration in the chamber
was 1.0 mg/mL. GUVs were electroformed at 65 °C using an AC signal
with a peak-to-peak amplitude of 3.0 V and a frequency of 10 Hz for
2 h (Siglent Technologies). GUVs were removed from the electroformation
chamber and diluted 1:1000 with MOPS buffer in an Attofluor imaging
chamber (Thermo Fisher Scientific). Prior to imaging, GUVs were exposed
to 1 μM EuTc and/or 10 nM CTxB-Alexa647. A minimum of 30 GUVs
among ≥3 individual preparations were examined for signal partitioning,
and representative images were chosen to produce in the figures.
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