Which step is described as part of the LCB procedure?

Which step is described as part of the LCB procedure?

• A. Gently heat after applying stain
• B. Special care in obtaining inoculum with needle
• C. Use large portion of medium with mycelium
• D. Do not apply coverslip

Answer: (B)

When preparing slides with lactophenol cotton blue, the key is to start with a tiny, clean sample from the culture. Special care in obtaining inoculum with a sterile needle ensures you pick a thin, well-distributed amount of fungal material rather than a chunk of agar. This allows the hyphae and other structures to be seen clearly after staining, because the smear remains thin enough for the stain to penetrate and for objective details to be visualized under the microscope. A large piece of medium or mycelium would crowd the slide and obscure features, making identification harder. Heating after staining isn’t a standard part of the protocol and could distort structures, while mounting with a coverslip is typically necessary to observe the sample properly.

When preparing slides with lactophenol cotton blue, the key is to start with a tiny, clean sample from the culture. Special care in obtaining inoculum with a sterile needle ensures you pick a thin, well-distributed amount of fungal material rather than a chunk of agar. This allows the hyphae and other structures to be seen clearly after staining, because the smear remains thin enough for the stain to penetrate and for objective details to be visualized under the microscope. A large piece of medium or mycelium would crowd the slide and obscure features, making identification harder. Heating after staining isn’t a standard part of the protocol and could distort structures, while mounting with a coverslip is typically necessary to observe the sample properly.