Collagenases (MMP-1,-8,-13) are enzymes belonging to the family of matrix metalloproteinases and play significant roles in tumor invasion and metastasis. Collagen gel zymography and in situ zymography are used in the analysis of collagenases. Existing in situmethods are not sufficient for in vitro determination of local activities of collagenases. In this study, it was aimed to develop the "cell in situ collagen zymography" method. 8505C anaplastic thyroid cancer cells were first fixed with zinc, methanol and ethanol, and the effects of fixatives on cell adhesion were investigated. Three different gel models were formed in a 96-well plate with gel mixtures containing type-1 collagen/ fluorescent-labeled type-1 collagen with different ratios. These models are: cell on bottom- gel mixture on top, gel mixture on bottom- cell on top and cell between two different gel mixtures (sandwich). The effects of the fixatives used on the fluorescence signal intensity were examined on the formed collagen gels and the sandwich model was determined as appropriate. In this model, the ratio of type-1 collagen: fluorescent-labeled type-1 collagen was applied as 2:1. We observed that thyroid cancer cell adhesion and fluorescence intensity increased with methanol fixation compared to zinc and ethanol fixation. In the sandwich model we developed, we observed that the cells and fluorescence signal were better preserved in the fixation based on protein precipitation between two thin collagen gel layers. Consequently, we developed the “cell in situ collagen zymography” method that can be used in vitro by adapting it to a 96-well plate.