J Assist Reprod Genet (2013) 30:1389–1393
DOI 10.1007/s10815-013-0064-4
TECHNOLOGICAL INNOVATIONS
The heat is on: room temperature affects laboratory
equipment–an observational study
Julia M. Butler & Jane E. Johnson & William R. Boone
Received: 14 May 2013 / Accepted: 17 July 2013 / Published online: 7 August 2013
# Springer Science+Business Media New York 2013
Abstract
Objective To evaluate the effect of ambient room temperature
on equipment typically used in in vitro fertilization (IVF).
Design We set the control temperature of the room to 20 °C
(+/−0.3) and used CIMScan probes to record temperatures of
the following equipment: six microscope heating stages, four
incubators, five slide warmers and three heating blocks. We
then increased the room temperature to 26 °C (+/−0.3) or
decreased it to 17 °C (+/−0.3) and monitored the same equip-
ment again. We wanted to determine what role, if any, changing
room temperature has on equipment temperature fluctuation.
Results There was a direct relationship between room tem-
perature and equipment temperature stability. When room
temperature increased or decreased, equipment temperature
reacted in a corresponding manner. Statistical differences be-
tween equipment were found when the room temperature
changed. What is also noteworthy is that temperature of equip-
ment responded within 5 min to a change in room temperature.
Conclusions Clearly, it is necessary to be aware of the affect
of room temperature on equipment when performing assisted
reproductive procedures. Room and equipment temperatures
should be monitored faithfully and adjusted as frequently as
needed, so that consistent culture conditions can be maintained.
If more stringent temperature control can be achieved, human
assisted reproduction success rates may improve.
This research was submitted to the Faculty of Eastern Virginia Medical
School in partial fulfillment of the requirement for the degree of Master of
Science in Biomedical Sciences-Clinical Embryology and Andrology.
Norfolk, Virginia May 2012.
Capsule Because modest changes in ambient air can affect surrounding
equipment, it is necessary to be aware of room temperature when
performing assisted reproductive procedures.
J. M. Butler (*) : J. E. Johnson : W. R. Boone
Department of Obstetrics and Gynecology, Greenville Health System
University Medical Group, 890 W. Faris Rd., Suite 470,
Greenville, SC, USA
e-mail: jbutler@ghs.org
Keywords Assisted reproductive technology . ART . Room
temperature . Equipment stability
Introduction
Temperature is an essential component of cellular physiology
and a critical aspect of embryo culture [1]. It is well docu-
mented in the literature that temperature stability is necessary
during in vitro fertilization (IVF) [2, 3]. In many species such
as rabbit [4], bovine [5, 6], mouse [3, 7], rat [8, 9] and human
[2, 3, 10–12] in vitro exposure to anomalous temperatures
affects gamete competency, embryo development, and off-
spring survival.
Because temperature can influence in vivo and in vitro
reproduction, temperature control is vital in assisted reproduc-
tion to ensure that no gametes or embryos are exposed to
extreme temperature fluctuations. Stringent temperature con-
trol is essential not only during embryo culture, but also
during manipulation of gametes and embryos [2, 3]. Incuba-
tors, microscope heating stages, slide warmers, and heating
blocks must all be calibrated and monitored regularly during
use [2, 3] to avoid unnecessary exposure of gametes and
embryos to detrimental conditions.
While IVF laboratories are more capable of strict
control over equipment temperature, room temperature
is not as manageable. Most laboratories’ room tempera-
tures fluctuate between 20 and 25 °C [13]. Room tem-
perature and ambient temperatures are directly affected
by the heating and cooling system. Due to air flow
patterns, hot and cold spots occur in the laboratory
[13]. These hot and cold spots, therefore, affect equip-
ment temperature as these airflow patterns will change
during heating and cooling cycles. While many scientists
believe surrounding air affects equipment temperature [14–18],
the literature is lacking to support this theory.
The aim of this study is twofold. First, we demonstrate the
effect that room temperature has on laboratory equipment, and
1390 J Assist Reprod Genet (2013) 30:1389–1393

second, we reveal how quickly a change in room temperature
can effect laboratory equipment. This narrowly focused study
pertaining to a specific aspect of ART labs will prompt other
labs to access their own laboratory environments.
Materials and methods
Background
This was an observational study that was conducted in the
assisted reproduction laboratory at Greenville Memorial Hos-
pital, located in Greenville, South Carolina from July 5 to July
26, 2011. Because patient data were not utilized, no Internal
Review Board was required.
Mechanism of action for CIMScan
taken with the NIST thermometers and these were averaged
(data not shown), to verify that CIMScan probes and NIST
thermometers all agreed within +/−1.0 °C.
Because heating elements operate in a sine wave fashion
[3], it was decided that +/−1.0 °C was the closest that could be
achieved at any particular point in time. This value was
selected because the NIST thermometers may be recording
the temperature at any point along the sine wave and may not
represent an average temperature over a 5 min interval as did
CIMScan.
As a result of the comparison between the NIST thermom-
eters and CIMScan, we decided agreement was comparable
and the rest of the measurements were made using CIMScan
only. Equipment included in the study is listed in Table 1.
Experiment I: effect of room temperature changes
on equipment temperatures

We used CIMScan (CIMTechniques, Inc., Beaufort SC) tech-
nology to monitor temperature changes in equipment.
CIMScan worked via wireless temperature probes attached
to each piece of monitored equipment. The probes sent mea-
surements to a monitoring station. These data were sent to a
server where it was stored and compared against user-assigned
alarm limits. If an alarm was detected, users were immediately
notified of the condition. Data were also delivered to the user
program where values were displayed.
Validation of CIMScan
The control temperature for the room was set at 20 °C
(+/−0.3 °C). The room temperature was increased to
26 °C (+/−0.3 °C) or decreased to 17 °C (+/−0.3 °C) and
equipment temperature was recorded. The room temperature
ran for three consecutive days at each temperature. Shields
were placed over air vents to divert direct airflow away from
equipment. When the room temperature was stable, the tem-
perature of each piece of equipment was recorded every 5 min
over a 12 h period for a total of 129 measurements. The
temperature variance (for each individual piece of equipment)
and the mean temperature variance (comparison between the

To ensure accuracy, CIMScan was validated with the use of

two manufacturer calibrated NIST Greisinger GMH 3230 Table 1 Manufacturer/model number of equipment used in an in vitro
digital thermometers (NIST; Greisinger Electronics, Germa- fertilization laboratory
ny). Before the study began, temperature was recorded for Equipment Manufacturer/model number
each piece of equipment. The two thermometers measured
room temperature and a randomly selected piece of equipment Heating block 3 Labline Multiblock/2052
to ensure that both thermometers agreed within +/−0.1 °C. Heating block 5 Barnstead International Modular Block/2003
The average difference between the two thermometers at room Slide warmer 1 Precision Scientific Inc./66632
temperature was 0.03 °C. The average difference between the Slide warmer 2 Labline/26020
two thermometers when measuring equipment was 0.04 °C. Slide warmer 3 Labline/26005
The p-value associated with room temperature was P=0.15 Slide warmer 4 Labline/26020
and the p-value associated with equipment was P=0.13. From Slide warmer 5 Labline/26020
this evaluation it was agreed that the two thermometers were Dry incubator 2 Boekel Scientific/132000
reading similarly and could be used to validate CIMScan. Incubator 1 Thermo Electron Corp, Forma series II/3140
So as not to interfere with equipment use, CIMScan probes Incubator 11 Thermo Electron Corp, Forma series II/3110
were attached towards the edges of work surfaces of the Incubator 12 Thermo Electron Corp, Forma series II/3110
equipment. Thirty measurements were taken for each piece Scope 1 Nikon/A0 stereomicroscope
of equipment at a specific point on its work surface using the Scope 2 Nikon/SMZ 1550
NIST thermometers. Once the values were averaged, they Scope 3 Nikon/Diaphot-MD
were compared to the values reported by CIMScan. A techni- Scope 4 Nikon/Diaphot-MD
cian re-calibrated the CIMScan probes to match the average Scope 5 Nikon/SMZ 1550
temperature of the work surface area of each piece of equipment. Scope 6 Eclipse/TES TE2000-S
After adjustments were made, another 30 temperatures were
J Assist Reprod Genet (2013) 30:1389–1393 1391

different types of equipment) were determined at the three
different room temperatures (17 °C vs. 20 °C and 20 °C vs.
26 °C).
Table 2 Effect of temperature on equipment at 20 °C vs. 26 °C. One
hundred and twenty-nine measurements were taken for each piece of
equipment. Measurements from identical pieces of equipment were
pooled for analysis

Equipment 20 °C 26 °C Paired P value

Experiment II: effect of sudden room temperature changes difference
on equipment temperatures
Scope 1 34.30±0.02 36.16±0.07 1.86±0.07 <0.0001
To determine how quickly equipment can respond to room Scope 6 35.00±0.07 37.05±0.16 2.05±0.17 <0.0001
temperature, a sudden temperature increase that occurred dur- Scope 3, 4 35.15±1.13 36.90±0.21 1.75±1.32 <0.0001
ing the investigatory period was accessed to observe the effect Scope 2, 5 33.08±0.79 35.02±0.36 1.94±0.46 <0.0001
that a rapid increase or decrease in room temperature had on Slide warmer 1 34.38±0.16 36.38±0.08 3.39±0.20 <0.0001
equipment. Slide warmer 3 32.74±0.36 35.89±0.47 3.14±0.55 <0.0001
Slide warmer 2, 4, 5 32.82±0.29 35.76±0.39 2.94±0.48 <0.0001
Experiment III: comparison of the temperature stability Heating block 3 34.70±0.93 35.19±0.90 0.49±1.21 <0.0001
of analog and digital microscope stage warmers Heating block 5 37.12±0.08 37.67±0.05 0.55±0.09 <0.0001
Dry incubator 2 36.79±0.17 37.7±0.16 0.88±0.27 <0.0001
A comparison of analog and digital microscope stage warmers Incubator 1 37.0±0.08 37.0±0 0.008±.09 0.32
was performed to determine if one type was more temperature Incubator 11 36.60±0.02 37.0±0.009 0.10±0.009 <0.0001
stable than the other. Three hundred and eighty seven temper- Incubator 12 36.4±0 36.6±0 0.20±0.0 <0.0001
ature measurements for three analog and 387 temperature
measurements for three digital stage warmers were taken at
room temperatures of 17 °C and 26 °C. The results for each
type of stage warmer at each room temperature were averaged 17 °C versus 20 °C
and compared.
There was a direct relationship between room temperature and
equipment temperature when comparing equipment tempera-
Statistical analysis
tures at 17 °C and 20 °C. Again, for all microscope stages,
slide warmers, heating blocks and incubators, a statistically
We attempted to take sufficient temperatures for each piece of
significant temperature difference was observed at a room
equipment with the hopes of reducing variability and bias.
temperature of 17 °C versus 20 °C (Table 3).
There were no changes to the study outcomes once the trial
commenced. No data were lost. All statistical analyses of the
data were performed using a Student’s paired t test. Statistical
significance was defined as P<0.05. Table 3 Effect of temperature on equipment at 17 °C vs. 20 °C. One
hundred and twenty-nine measurements were taken for each piece of
equipment. Measurements from identical pieces of equipment were
pooled for analysis
Results
Equipment 17 °C 20 °C Paired P value
difference
Experiment I: effect of gradual room temperature changes
on equipment temperatures Scope 1 33.14±0.08 34.3±0.02 1.15±0.08 <0.0001
Scope 6 33.69±0.09 35.0±0.07 1.30±0.11 <0.0001
20 °C versus 26 °C Scope 3, 4 32.60±0.43 35.10±1.13 2.50±0.77 <0.0001
Scope 2, 5 30.05±0.09 33.08±0.79 2.50±0.73 <0.0001
Equipment temperature was compared at two different room Slide warmer 1 28.01±0.17 30.45±0.08 2.44±0.18 <0.0001
temperatures to investigate the affect of room temperature on Slide warmer 3 30.54±0.42 32.74±0.36 2.20±0.53 <0.0001
equipment stability (Table 2). Measurements from identical Slide warmer 2, 4, 5 30.32±0.25 32.82±0.29 2.50±0.39 <0.0001
models of microscope stages and slide warmers were pooled Heating block 3 34.15±0.87 34.70±0.93 0.55±1.32 <0.0001
for analysis. When comparing equipment temperature at Heating block 5 36.71±0.03 37.12±0.03 0.41±0.05 <0.0001
20 °C and 26 °C, there were statistically significant tempera- Dry incubator 2 36.27±0.12 36.78±0.12 0.50±0.18 <0.0001
ture differences between all microscope stages, slide warmers, Incubator 1 36.90±0.01 37.00±0 0.10±0.01 <0.0001
heating blocks and two of the three incubators, regardless of Incubator 11 36.50±0 36.60±0 0.10±0 <0.0001
model. The only piece of equipment that maintained the set Incubator 12 36.40±0 36.30±0 0.10±0 <0.0001
temperature was a non-humidified Forma incubator.
1392
Experiment II: effect of sudden room temperature changes
on equipment temperatures
Equipment temperature responded within 5 min to a sudden
change in room temperature (Fig. 1). Data demonstrated a
direct relationship between a sudden upward spike in room
temperature and an increase in equipment temperature.
Experiment III: comparison of the temperature stability
of analog and digital microscope stage warmers
A comparison between digital and analog equipment was
performed. Three analog and three digital microscope stage
warmers were compared at 17 °C and 26 °C. Analog micro-
scope stage warmer temperatures ranged from 30.3 °C to
33.4 °C at 17 °C and 36.1 °C to 37.4 °C at 26 °C. Digital
microscope stage warmer temperatures ranged from 30.3 °C
to 33.9 °C at 17 °C and 34.2 °C to 37.3 °C at 26 °C. Both
analog and digital microscope stage warmers were similar in
their ability to maintain temperature at 17 °C (P=0.35). A
statistical difference in their ability to maintain set temperature
was observed at 26 °C (P<0.0001).
Discussion
To the authors’ knowledge, this is the first time that a study
clearly demonstrates a direct relationship between room tem-
perature and laboratory equipment temperature. As room tem-
perature increased, the temperature of heating blocks, micro-
scope stage warmers, slide warmers and incubators increased
regardless of make or model (P<0.0001). The same equip-
ment decreased in temperature as room temperature decreased
(P<0.0001). Microscope stage warmers and slide warmers
had the greatest temperature fluctuation compared to heating
Fig. 1 Direct effect of a sudden
upward spike in room
temperature on equipment. Room
temperature is shown in red and
microscope stage temperature is
shown in blue
J Assist Reprod Genet (2013) 30:1389–1393
blocks and incubators. Heating blocks and incubators did have
some temperature fluctuation; however, these slight tempera-
ture fluctuations most likely do not affect gametes and em-
bryos. This study also demonstrates that equipment tempera-
ture reacts rapidly to sudden changes in room temperature. If
room temperature spikes, equipment temperature will show
the same increase within 5 min.
Since digital heating elements are a more recent technolo-
gy, there may be a bias that digital equipment maintains
temperature more consistently than analog equipment; how-
ever, this was not demonstrated in this study. When comparing
digital and analog microscope stage warmers, there was no
statistical difference between the two at 17 °C. However,
when the room temperature increased to 26 °C, there was a
significant temperature difference between the digital and
analog stage warmers. Analog stage warmers were able to
hold closer to the set point at both temperatures.
Though there is limited clinical correlation in this study,
there is an implication that changes in lab equipment temper-
ature may have an effect on gamete and embryo development.
Equipment tested in our laboratory is certainly not an exhaus-
tive list of equipment used to maintain temperature of gametes
and embryos during ART procedures. Other examples include
different types of test tube warmers used during retrievals
[14], IVF workstations [18], and heated microscope objectives
[11, 15]. Each laboratory must evaluate their own equipment
and optimize temperature control.
For ethical reasons, it is not possible to assess the effect of
changes in room temperature on human embryos and gametes.
However, with the use of animal models, future studies in-
clude assessing changes in embryos and gametes as room
temperature changes.
The concept of room temperature fluctuation and its effect
on equipment temperature needs to be taken into consider-
ation when constructing an ART laboratory. Such a facility
J Assist Reprod Genet (2013) 30:1389–1393
should not be placed so as to have the walls be part of the
exterior of a building. A facility should not reside on the top
floor of a building where outside temperatures that change
with the seasons of the year have the potential of altering the
inside room temperatures. The use of windows in the walls of
an ART laboratory should be assessed carefully due to the
transfer of heat and cold through glass.
In a field where temperature is critical, it is troublesome to
think that our equipment is quite unstable. During the perfor-
mance of assisted reproduction procedures, there are many
instances when gametes and embryos can be exposed to
temperature fluctuations (i.e., oocyte retrieval and embryo
transfer, incubator door openings, changes in location from
incubator to microscope stage). These fluctuations are magni-
fied if the room temperature is not constant. Perhaps room
temperature is one of the culprits in the inefficiency of human
assisted reproduction.
Conclusion
Clearly, it is necessary to be aware of the affect of room
temperature on equipment when performing assisted reproduc-
tion procedures. Room and equipment temperatures should be
monitored faithfully and adjusted as frequently as needed, so
that consistent culture conditions can be maintained. If more
stringent temperature control can be achieved, human assisted
reproduction success rates may improve.
Acknowledgments The authors' would like to acknowledge H. Lee
Higdon III, Ph.D. for his statistical assistance.
Conflicts of interest None of the authors have a conflict of interest.
All funding for this research came from within the department.
References
1. Ye J, Coleman J, Hunter MG, Craigon J, Campbell KHS, Luck MR.
Physiological temperature variants and culture media modify meiotic
progression and developmental potential of pig oocytes in vitro.
Reproduction. 2007;133:877–86.
1393
2. Boone WR, Higdon III HL, Johnson JE. Quality management issues in
the assisted reproduction laboratory. J Reprod Stem Cell Biotechnol.
2010;1:30–107.
3. Lane M, Mitchell M, Cashman KS, Feil D, Wakefield S, Zander-Fox
DL. To QC or not to QC: the key to a consistent laboratory? Reprod
Fertil Dev. 2008;20:23–32.
4. Ulberg LC, Burfening PJ. Embryo death resulting from adverse
environment on spermatozoa or ova. J Anim Sci. 1967;26:571–7.
5. Aman RR, Parks JE. Effects of cooling and rewarming on the meiotic
spindle and chromosomes of in vitro-matured bovine oocytes. Biol
Reprod. 1994;50:103–10.
6. Ju JC, Jiang S, Tseng JK, Parks JE, Yang XZ. Heat shock reduces
developmental competence and alters spindle configuration of bovine
oocytes. Theriogenology. 2005;64:1677–89.
7. Pickering SJ, Johnson MH. The influence of cooling on the organi-
zation of the meiotic spindle of the mouse oocyte. Hum Reprod.
1987;2:207–16.
8. Almeida PA, Bolton VN. The effect of temperature fluctuations on
the cytoskeletal organization and chromosomal constitution of the
human oocyte. Zygote. 1995;3:357–65.
9. Cockroft D, New D. Abnormalities induced in cultured rat embryos
by hyperthermia. Teratology. 1978;17:277–81.
10. Kimmel GL, Williams PL, Claggett TW, Kimmel CA. Response-
surface analysis of exposure-duration relationships: the effects of
hyperthermia on embryonic development of the rat in vitro. Toxicol
Sci. 2002;69:391–9.
11. Wang WH, Meng L, Hackett RJ, Oldenbourg R, Keefe MD. Rigor-
ous thermal control during intracytoplasmic sperm injection stabilizes
the meiotic spindle and improves fertilization and pregnancy rates.
Fertil Steril. 2002;77(6):1274–7.
12. Langley MT, Marek MA, Doody KM, Nackley AC, Doody KJ.
Importance of micro-drop temperature measurement. Embryol
Newsl. 2003;6:3–5.
13. Bove R. Temperature measurement in the clinical laboratory. Good
isn’t good enough. Med Lab Obs. 2011;43:36–9.
14. Yeung QSY, Briton-Jones CM, Tjer GCC, Chiu TTY, Haines C. The
efficacy of test tube warming devices used during oocyte retrieval for
IVF. J Assist Reprod Genet. 2004;21:355–60.
15. Cooke S, Tyler JPP, Driscoll G. Objective assessments of temperature
maintenance using in vitro culture techniques. J Assist Reprod Genet.
2002;19:368–75.
16. Conaghan J, Steel T. Real-Time pH profiling of IVF culture medium
using an incubator device with continuous monitoring. J Clin
Embryol. 2008;11:15–6.
17. Khabani A, Tufts K, Craig L, Soules M, Scott L. Cooling and
warming rates in microdrops for embryo culture. Fertil Steril.
2003;80:S929-3.
18. Swain JE, Cabrera L, Xu X, Smith GD. Microdrop preparation
factors influence culture-media osmolality, which can impair mouse
embryo preimplantation development. Reprod Biomed Online.
2012;24:142–7.