Abstract
There are balances sensitive enough to measure the masses of small arthropod muscles, but preparing and handling these tissues can be problematic. It is often difficult to remove intact muscles and, once removed, they are easily damaged. The method described in this paper can be used to determine the mass of free muscle tissue, muscles that are left attached to pieces of the exoskeleton for easier handling, and muscles within intact arthropod appendages. This method employs a spectrophotometer to measure the amount ofdye released when stained muscle tissue is dissolved in sodium hydroxide. A number of biological stains may prove useful in this technique, although they must be able to withstand an extremely high pH. The example presented below uses Wilkey’s doublestain, purchased from Arthropod Slidemounts, P.O. Box 185, Bluffton, IN 46714. This stain contains 5 ml of5% aqueous acid fuchsin and 10 ml of5% aqueous lignin pink in 100 ml of modified Essig’s aphid clearing fluid (Essig 1948), consisting of 20 parts 85% lactic acid, 2 parts liquified phenol, 4 parts glacial acetic acid, and part distilled water. To test this procedure, I determined the first leg muscle masses for a developmental series of the spider Miagrammopespinopus Chickering (Uloboridae) from the U.S. Virgin Islands (Opell 1987). These spiders’ live weights ranged from 0.63 to 4.27 mg. I killed and preserved them in 75% ethanol, removed their two first legs at the trochanter-femur joint, and stained these legs for 20 days at room temperature in Wilkey’s stain. This long staining time assured that the muscles ofeven the largest legs were fully saturated with stain. To remove stain trapped within hemolymph channels, I then rinsed legs in five changes of 75% ethanol for one day each. I next transferred them to 0.7 ml of 5% (weight/volume) NaOH and digested them at 70C for 90 minutes.
Highlights
There are balances sensitive enough to measure the masses of small arthropod muscles, but preparing and handling these tissues can be problematic
To establish a calibration curve (Fig. 2) for muscle mass, I dissected out the leg muscles of another uloborid spider, Octonoba sinensis (Simon)
When their masses were plotted against their transmittance to 480 nm light (Fig. 2), the resulting regression is highly significant (p
Summary
There are balances sensitive enough to measure the masses of small arthropod muscles, but preparing and handling these tissues can be problematic. I determined the first leg muscle masses for a developmental series of the spider Miagrammopes pinopus Chickering (Uloboridae) from the U.S Virgin Islands (Opell 1987). As Figure illustrates, this is the wavelength of maximum absorbance for Wilkey’s stain in a 5% NaOH solution.
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