Temperature control for microscope imaging: incubation box

Introduction

Following biological processes using microscopy imaging requires maintaining the cells as healthy as possible and sometimes for very long period for time depending on the manipulator experiments. Several temperature controllers are available; they all have positive and negative features, it is lends on microscope users to evaluate which one is best suited for their experiments. Incubator box is a stable way to control the cells temperature and environment.

Precision-30x28How does it work?

Incubator box is a plexiglas box, which is fixed on and covers most of the microscope. It provides a multi-parameters (gas, pH, humidity and temperature) controlled environment. Notably, it allows the whole thermalization of the microscope with electric heating or flow-induced heating and cooling. This system is particularly suitable for long-term imaging as it maintains the temperature of the microscope to ensure a steady focus level during acquisition.

Precision-30x28Advantages

Homogeneous sample temperature

Because the whole microscope is thermalized, this system does not generate a strong thermal gradient on the sample, thus limitating possible artifacts.

High resolution microscopy

Oil or water immersion objectives do not generate thermal gradients since tey are at the same température as the sample. Thus, contrary to microscope stage heaters, it is not requires to add an objective collar. However, the objectives should withstand the temperature settings of the incubation box (thermal expansion).

For long-term experiments

Incubation box control the whole cell environment and is thus well-suited for long term experiments and time-lapse imaging. This system can be use with Petri dishes, chamber slides or culture-cell flasks.

Easy to use

Once the box is set-up, it is easy to use, and requires no particular training from the user.

Precision-30x28Disadvantages

Speed of the temperature switch (30 min)

The incubation box requires the thermalization of the whole microscope leading to slow temperature changes due to the system inertia. Thus, with this system, it is not possible to use temperature to study dynamic biological processes. Also, incubation box system has to be turn on 1 or 2 hours before the beginning of the experiments for equilibration purposes.

Temperature range

Ambient+3°C to 50°C, except for some model of incubation boxes integrating air conditioning systems which enable to decrease temperature below ambient.

Mono temperature control

With this system only one temperature can be pre-set-up. While other systems allows to set two temperature at once and shift from one to the other ultra-rapidly.

Lack of active cooling system

Since incubation boxes do not enable direct cooling of the sample, strong temperature increases can occur when turning on DIC illumination or during extended observation under epi-fluorescence microscopy.

Requires a microscope chamber top

Because the volume of the incubation is large, for sample CO2 regulation, it is recommended to place the dish or plate inside a microscope chamber top.

 

CherryTemp heater/cooler for live-cell imaging

Dynamic & fast : 10 seconds temperature shifts


Stable & Precise : Long term stability


Discover the C. elegans heater/cooler

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