To create cavities of various thicknesses, we used spacers, 1 mm thick (Press-to-seal Silicone Isolator, P24744, Invitrogen) and 120 μm thick (SecureSeal Imaging spacer, GBL654008, Sigma-Aldrich). The 120 μm spacers have been stacked to gradually vary the thickness from 120 μm to 840 μm. The same volume (7 μL – OD 0.08) of bacterial suspension was deposited in each seal. Each well was then completely filled with the appropriate culture media, and hermetically sealed with a glass coverslip. After a sedimentation time (from 1 h 30 min to 20 h depending on the strain and on the well thickness), the samples were incubated at the appropriate growth temperature without shaking. Images of the sample were taken just before incubation (0 h) and then every hour with a CMOS camera (DCC1545M-GL – Thorlabs) plugged to an inverted phase-contrast microscope (Eclipse TS2 – Nikon, 10× objective). A mark was placed on each sample to observe the same area throughout the experiment. Only Geobacillus stearothermophilus and Thermus thermophilus were studied this way.
Secureseal imaging spacer
Manufactured by Merck Group
Sourced in Canada
The SecureSeal Imaging spacer is a laboratory equipment product designed to create a secure and controlled environment for various imaging applications. It serves as a physical barrier to isolate and protect samples during imaging procedures. The core function of the SecureSeal Imaging spacer is to provide a sealed and stable platform for imaging tasks, ensuring the integrity and consistency of the imaging process.
Automatically generated - may contain errors
2 protocols using secureseal imaging spacer
1
Bacterial Growth Monitoring at Various Depths
2
Lipid Droplet Quantification in Morula Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For lipid droplets quantification, morula-stage embryos were preferred and selected over blastocysts because the absence of fluid-filled cavity allows a better imaging resolution. A first staining of morula embryos was performed using 300 nM MitoTracker Orange CMTMRox (ThermoFisher Scientific, Burlington, ON, Canada) in a flushing medium (ViGRO, Bioniche, Belleville, ON, Canada) at 37°C during 30 min. Then, a second staining was performed using 3 μg/mL Bodipy FL (ThermoFisher Scientific) and 1 μg/mL Hoechst blue dye 33342 (ThermoFisher Scientific) diluted in flushing medium and incubated during 10 min at 37°C. Finally, embryos were fixed in 4% paraformaldehyde (Sigma-Aldrich, Oakville, ON, Canada) for 15 min at room temperature, washed 3 times in flushing medium, and mounted on a microscope slide using secure Seal imaging spacer (Sigma-Aldrich).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!