| Early- to mid-stage egg chambers dissected from well-fed females |
Halocarbon oil (series 95) |
Avoids dehydration/hypoxia |
|
|
Early stages: problem of activation and loss of MT organization in aqueous media |
|
|
Oil has higher RI than water |
|
|
Good environment for injection |
|
|
|
| Late-stage egg chambers dissected from well-fed females |
Grace’s medium (Sigma) |
Provides ionic and osmotic balance and nutrients |
|
|
Late stages not susceptible to problems of early stages |
|
|
|
| Embryos, dechorionated and dehydrated |
Halocarbon oil (series 700) |
RI similar to glycerol |
|
|
Halocarbon oil prevents excess dehydration while avoiding hypoxia (which causes changes to the cell cycle) |
|
|
Good environment for injection |
|
Halocarbon oil with breathable Teflon membrane |
Better dehydration prevention for long-term development studies |
|
|
Better bright-field imaging |
|
|
Can help squash specimen for greater optical clarity (reduced spherical aberration) |
|
Aqueous medium |
Can use WI objectives, which have longer working distance with high NA |
|
|
|
| Spread macrophages from third instar larvae |
Culture-slide-mounted coverslips treated with ConA in a humidified overchamber |
Inverted microscope, 100X 1.4-NA oil objective |
|
Six-well tissue-culture plate |
Upright microscope, 60X 0.9-NA dipping objective |
|
Schneider’s insect medium with 5% FCSb |
|
|
|
|
| Neuronal cell cultures from larval brains, overnight culture |
Dissociation by enzyme cocktail (Kraft et al. 1998) |
Inverted microscope, 100X 1.4-NA oil objective |
|
Schneider’s insect medium with 5% FCS |
|
|
Coverslips treated with ConA (16 μg/mL) and laminin (5 μg/mL) |
|
|
|
|
| Whole larval fillet |
Schneider’s insect medium with 5% FCS |
Upright microscope, 60X 0.9-NA dipping objective |
|
Sylgard mounting chamber |
|
|
| aAbbreviations: MT: microtubule; RI: refractive index; WI: water immersion; FCS: fetal calf serum; NA: numerical aperture.
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| bFrom Sigma-Aldrich.
|
