Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation

Materials and Reagents

1. Protective gear (gloves, goggles, masks, lab coat)

2. Deionized water

3. Sodium pentobarbital (Sigma, catalog number: P-010 )

4. 1.5% sodium pentobarbital (in saline)

5. NaCl (Sigma-Aldrich, catalog number: S7653 )

6. Saline (0.9% sodium chloride)

7. NaH₂PO₄·2H₂O (Sigma-Aldrich, catalog number: 71645 )

8. Na₂HPO₄·12H₂O (Sigma-Aldrich, catalog number: 71663 )

9. Paraformaldehyde (PFA) (Sigma-Aldrich, catalog number: P6148 )

10. NaOH (Sigma-Aldrich, catalog number: S8045 )

11. Hydrochloric acid (HCl) (Sigma, catalog number: H1758 )

12. Heparin (Sigma, catalog number: H3149 )

13. Sodium azide (Sigma, catalog number: S2002 )

14. 4% paraformaldehyde (PFA, in 0.1 M Phosphate Buffer, pH 7.0-7.4) (see Recipes)

Equipment

1. Chemical fume hood

2. Dissecting forceps (RWD, catalog number: F12018-13 ) (Figure 1)

3. Ophthalmic forceps (RWD, catalog number: F12006-10 ) (Figure 1)

4. Hemostatic forceps (RWD, catalog number: F21002-12 ) (Figure 1)

5. Tissue scissors (RWD, catalog number: S13052-12 ) (Figure 1)

6. Fine scissors (RWD, catalog number: S12005-10 ) (Figure 1)

7. Vascular clamp (RWD, catalog number: R33001-48 ) (Figure 1)

8. Micro spatula (Thermo Fisher, catalog number: 21-101-10 ) (Figure 1)

9. 1 ml and 20 ml syringe (Figure 1)

10. 25- or 27-gauge infusion needle (Figure 1)

11. (Optional) Peristaltic pump (LONGERPUMP, catalog number: BT100-2J )

12. 2.5 L and 5 L beakers

13. Thermometer (measuring range:100 °C, division value:1 °C)

14. Measuring cylinder

15. Analytical balance (SARTORIUS, catalog number: BSA124S )

16. Benchtop vacuum

17. 0.22 μm PVDF filter set (Nalgene, catalog number: 291-4520 )

18. 50 ml conical tube (Thermo Fisher, catalog number: 339652 )

19. Fume hood

20. Microwave

21. Magnetic stirrer with heating device (SCILOGEX, catalog number: MS7-H550-Pro )

Figure 1. Surgical instruments

Procedure