Temperature control of C. elegans samples during live-cell imaging
We developped the world’s fastest heater-cooler for microscopy. It enables ultra-fast temperature shifts [10 seconds] from any temperature to any other in the 5-45°C range. CherryTemp fits uprights ad inverted microscopes. You can now imùage and control the temperature of your favorite C.elegans.
Induce entry or exit of “Dauer” stage
Heat shock: control the timing of temperature exposure
Used for embryos or adult worm studies.
Follow GFP-tagged mutants and the phenotype induction
C. elegans thermosensitive embryo mutants (CYK-4 ts) examined under the microscope with a CherryTemp platform.
Application example: C. elegans embryogenesis control by CherryTemp
During our experimentation, C. elegans thermosensitive embryo mutants (CYK-4 ts) were examined under the microscope with a CherryTemp platform. This system allowed to selectively modify the temperature of embryo samples between 16°C and 25°C resulting in cytokinesis arrest. This outlined unprecedently observed phenomenon during cell division.
Timelapse images of the 1st division of C. elegans embryo movie:
Divisions were recorded using a microscope (DM6000; Leica) equipped with epifluorescence and DIC optics and a camera (DFC 360 FX; Leica). Images were collected every 15 sec using a 63×/1.4 NA objective and LAS AF software (Leica). Embryos were imaged at 22°C and the temperature was shifted to 5°C at 0sec thanks to the special control stage equipment. After 195 sec at 5°C, the temperature was shifted back to 22°C. Temperature control and tempertaure shifts were performed using Cherry Temp. more information about this application note
Mounting embryos on CherryTemp (for agarose pad or Hanging Drop technique) :
Traditionally, C. elegans embryos have been imaged on standard microscope slides with a 2% agarose pad and a coverslip. This has the advantage of being easy to assemble and requiring no special equipment. Moreover, it is possibled with the use of an eyelash tool or ultrafine deer hair to group embryos together, allowing simultaneous imaging of multiple embryos. Additionally, the process of sandwiching the embryo between the coverslip and the agarose pad results in a slight compression of the embryos, which can improve imaging of fluorescently tagged proteins.
An alternate method termed “hanging drop” traps the embryos in a drop of M9 buffer held between the slide and coverslip with a Vaseline petroleum jelly spacer. This prevents compression and results in the embryos being more randomly oriented relative to the plane of the slide. This inconsistency in orientation ensures, at least in a subset of the embryos, that imaging occurs at the optimum viewing angle for most cells during embryonic development.
- Place a coverslip on the Cherry insert (24_ 60 mm, No. 1.5). Dissect worms in a watch glass and transfer embryos on the coverslip using a mouth pipette (or dissect directly on the coverslip). Place the provided spacer on the coverslip. The spacer will prevent embryo compression by the Cherry chip and will avoid evaporation of the sample buffer.
- Place the Cherry insert with the coverslip on the microscope stage adaptor. Install the Cherry chip on top of the spacer so that the drop of medium that contains the embryos is sandwiched between the imaging coverslip and the Cherry chip. Lock the chip in place using the two slide clasps (not shown).
- Using the Cherry software interface, control the temperature by activating the flow from one heat exchanger to the other (fast shift) or by entering a new set point for the currently flowing heat exchanger (slower shift) as desire throughout the experiment.
T. Davies, S. Sundaramoorthy, S.N. Jordan,M. Shirasu-Hiza, J. Dumont and J.C. Canman, 2016, “Using fast-acting temperature-sensitive mutants to study cell division in Caenorhabditis elegans“, Methods in Cell Biology, Volume 137, ISSN 0091-679X, http://dx.doi.org/10.1016/bs.mcb.2016.05.004
C. elegans imaging & temperature
Play with ts mutants from 15 to 26°C and more!
EASY EMBRYO DISSECTION
Directly dissect and place embryos on our dedicated device!
ULTRAFAST SHIFTS | 10 SECONDS
Shift in less than 10 seconds and observe restrictive temperatures on live
STABLE & PRECISE | 0.1°C
Fine-tune permissive and restrictive temperatures
FITS ANY MICROSCOPE
CherryTemp can be easily installed and shared between different microscopes. We have a set of ergonomic elements fitting main brands (Nikon, Zeiss, Olympus, Leica) for both upright and inverted scopes.
BELOW AMBIANT | 5 TO 45°C
COULD IT WORK FOR ME?
No shear stress
Our thermalization technology does not interfere with your cells. They remain happy in a standard two-cover-slips sandwich.
Thanks to our spacer, you can add medium and avoid evaporation. In addition, we have a closed fluidic circuit for a long-term and stable thermalization of your sample.
Ready in 30 seconds
This mounting system is plug-and-play and respects microscopes standards. Just focus on your experimentation!
No interference with your set-up
Your microscope set-up can already be complex this is why we’ve designed CherryTemp to ensure a smooth integration.
Fits any inverted microscope
You might want to use different microscopes, this is why we’ve designed CherryTemp to be compatible with any inverted microscope.
First installation in 20 minutes
You want to focus on your experimentation this is why we’ve made CherryTemp easy to install. CherryTemp is plug and play and comes along with a didactic installation guide.