27 Louise Hyslop Incubators have a critical role in providing a stable environment for optimal embryo development within the in vitro fertilization (IVF) laboratory. To this end, incubators regulate the temperature, gas concentration, and humidity. Initially, IVF laboratories only had the option of large single chamber incubators designed for larger scale tissue culture. Nowadays there is an increasing range of benchtop and time‐lapse incubators designed specifically for gamete and embryo culture. Closed systems for gamete and embryo culture which also facilitate manipulation have been developed as an alternative to stand‐alone incubators and IVF workstations. This chapter will outline the main features to consider when selecting between the varieties of incubators available on the market. Single chamber incubators were designed for large scale tissue culture with flasks rather than the small dishes used in IVF laboratories for gamete and embryo culture. Nevertheless, this style of incubator is currently the most widely used in UK IVF laboratories (Bolton et al. 2014). These incubators are available in a wide range of chamber volumes from approximately 20 to 200 litres (Figure 27.1). Figure 27.1 Example of (a) a large volume single chamber incubator with (b) segmented inner door to reduce gas loss in a single chamber incubator. The large capacity for multiple patients also makes them a popular choice as a back‐up incubator and/or a separate location for equilibration of culture dishes. It is not within the scope of this chapter to compare manufacturers and incubator models. The main incubator features which regulate the gas concentration, temperature, and humidity will be discussed with the advantages and disadvantages of each. Benchtop incubators have been designed specifically for the culture of gametes and embryos in IVF laboratories. An increasing number of IVF laboratories have introduced them for embryo culture but usually in combination with another incubator type (Bolton et al. 2014). These incubators usually have more than one chamber with a volume of less than 1 litre (Figure 27.2). Figure 27.2 Example of a benchtop incubator. Reproduced with permission of Cook Medical, Bloomington, Indiana, USA. The small chamber size restricts the maximum number of patient dishes but the smaller incubator footprint allows laboratories to have more than one benchtop incubator in the same space required for a single large chamber incubator. Standard morphology assessment provides only snapshots at specific time points during the dynamic embryo development process. Time‐lapse systems take digital images of embryos at set intervals and can provide information that can be missed with standard morphological assessment, such as the presence of multinucleation. Additional features provided by time‐lapse imaging include timing of cell divisions, cell division patterns, and intervals between cleavage cycles (Figure 27.3). Figure 27.3 Embryoscope™ time‐lapse system. Reproduced with permission of Vitrolife, Göteburg, Sweden. Two different types of time‐lapse system have been developed: Currently, there are five time‐lapse systems available (Table 27.1). Table 27.1 Comparison of commercially available time‐lapse systems.
Incubators in the Assisted Reproductive Technology Laboratory
Introduction
Incubator Types
Single Chamber Incubators
Benchtop Incubators
Time‐lapse Incubators
Primo Vision™
EEVA™
Embryoscope™
Miri TL®
Geri
Type of system
Standalone camera that can be placed in standard incubator
Standalone camera that can be placed in standard incubator
Combined camera/incubator
Combined camera/incubator
Combined camera/incubator
Incubator design
User defined
User defined
Single chamber incubator for up to six Embryoslides®
Benchtop incubator with six separate chambers
Benchtop incubator with six separate chambers
Illumination
Bright field, green LED (550 nm)
Dark field, red LED (625 nm)
Bright field, red LED (635 nm)
Bright field, red LED (635 nm)
Bright field, orange LED (591n m)
Maximum number of focal planes
11
1
17 but reduced depending upon frequency of image capture
7
11
Frequency of image capture
From 5 minutes
5 minutes
From 10 minutes
5 minutes
5 minutes
Maximum number of embryos per dish
16 – Primo Vision embryo culture dish
12 – EEVA dish
12 ‐ Embryoslide®
14 – CultureCoin culture dish
16 – Geri dish
Embryo culture
Group
Group
Single
Single
Single
Total number of embryos monitored
96
48
72
84
96
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