Related Experiment Videos
Fluorophore toxicity in mouse eggs and zygotes
K P Phillips1, W L Zhou, J M Baltz
1Ottawa Civic Hospital, Ontario, Canada.
Zygote (Cambridge, England)
|October 15, 1998
Summary
This study assessed fluorophore toxicity in mouse eggs using in vitro fertilization. MQAE demonstrated significant toxicity, while BCECF, SNARF-1, and Fura-2 showed no adverse effects on IVF or zygote membrane properties.
Area of Science:
- Cell biology
- Reproductive toxicology
- Biochemistry
Background:
- Ion-sensitive fluorophores are crucial for measuring intracellular ion concentrations.
- Fluorophore loading and excitation can generate toxic byproducts, raising concerns for mammalian egg and embryo research.
- Limited data exists on fluorophore toxicity in mammalian oocytes and early embryos.
Purpose of the Study:
- To evaluate the toxicity of commonly used ion-sensitive fluorophores in mouse eggs and zygotes.
- To assess potential detrimental effects associated with fluorophore loading and fluorescence excitation.
- To identify fluorophores safe for use in mammalian reproductive cell research.
Main Methods:
- In vitro fertilization (IVF) was used to assess fluorophore toxicity on mouse egg viability and fertilization success.
- Whole-cell patch-clamp recordings were employed to detect membrane damage in zygotes exposed to fluorophores.
- Four fluorophores (SNARF-1, BCECF, Fura-2, and MQAE) were tested, targeting pH, Ca2+, and Cl-.
Main Results:
- Cleavage of acetoxymethyl (AM) ester groups alone did not impact IVF success or zygote membrane electrical properties.
- Intracellular loading and subsequent fluorescence excitation of BCECF, SNARF-1, and Fura-2 were non-toxic under tested conditions.
- MQAE exhibited significant toxicity, both when used alone and when subjected to fluorescence excitation.
Conclusions:
- BCECF, SNARF-1, and Fura-2 are safe for quantitative ion measurements in mouse eggs and zygotes.
- MQAE poses a significant toxicity risk and should be used with caution or avoided in studies involving mammalian oocytes and embryos.
- These findings are critical for selecting appropriate fluorophores in reproductive biology research to ensure cell viability and accurate measurements.