I have experimented to assess the T-cell memory status in mice models. However, I am unable to separate the CD3+ T-cell cluster from the splenic cells using different anti-CD antibody markers, specifically CD3 (Picture attached here). I followed the standard protocol for sample preparation and staining except used a 1 mL pipette-tip instead of a syringe with a 22G needle to make a single cell suspension. Now my question is how can I solve this problem? Thank you in advance for your consideration and valuable suggestions.