Which acid-fast staining method does NOT use heat?

The Kinyoun method is referred to as a cold method for acid-fast staining, meaning it does not require heat to drive the stain into the cells. This is significant because heating can sometimes cause distortion of cell morphology; therefore, the Kinyoun method uses a more concentrated primary stain, which allows for effective staining without the need for heating. This method is particularly useful in clinical microbiology, especially for identifying Mycobacterium species, which are acid-fast bacilli.

In contrast, the Ziehl-Neelsen method involves heating the slide to promote the uptake of the stain, which is particularly important for some methods of staining organisms that possess a thick waxy cell wall. The Schaeffer-Fulton method pertains specifically to spore staining, which also involves the application of heat to facilitate the staining of bacterial spores. Gram staining, on the other hand, is a separate method entirely that classifies bacteria based on their cell wall properties and does not involve the acid-fast principle. Thus, Kinyoun's use of a cold staining technique sets it apart from the other methods mentioned.