Mirxes Holding Company Limited · HKEX:2629
Mirxes — Technical Presentation (April 2025)
14 slides from Mirxes's technical presentation, Apr 7, 2025
Slide 1: Stereo-seq Sample Preparation Guidelines
mirxes TO KNOW. TO ACT. Stereo-seq Sample Preparation Guidelines Apr 2025 Document No.: MGL-F-504-02 Version No.: 4.0 Effective Date: 7 Apr 2025
Slide 2: General instructions and applicable chip sizes
• Instructions provided in this manual are intended for general use only and optimization may be required for specific applications. • Instructions provided is applicable to 1cm*1cm, 1cm*2cm, 2cm*2cm and 2cm*3cm Stereo-seq chip sizes. Please refer to section 1 and section 2 for specific information
Slide 3: Sample requirements and sectioning plane size
- The sectioning plane should not exceed (refer to the figures on the right) • 0.9 cm x 0.9 cm x 1cm (for 1cm*1cm Stereo-seq Chip) • 0.8 cm x 1.8 cm x 1 cm (for 1cm*2cm Stereo-seq Chip) • 1.8 cm x 1.8 cm x 1 cm (for 2cm*2cm Stereo-seq Chip) • 1.8 cm x 2.7 cm x 1 cm (for 2cm*3cm Stereo-seq Chip) - Ap
Slide 4: 2.1 Prepare these recommended apparatuses/materials in advance
2.1 Prepare these recommended apparatuses/materials in advance
Slide 5: 2.1 Prepare apparatuses/materials in advance (continued)
2.1 Prepare these recommended apparatuses/materials in advance (continued) 1. A box of crushed ice and pre-cool OCT on ice for 10 min in advance. 5. A box of dry ice. 2. 2 pieces of stainless-steel base molds slightly larger than the tissue 6. A metal block that has a flat surface to support the sta
Slide 6: 2.2 Sample embedding steps
2.2 Sample embedding steps 1. Upon harvesting within 30 min, use sterile gauze or dust free paper to 2. Place the tissue in pre-cooled OCT and wrap the tissue evenly with OCT using absorb excess liquid on the tissue surface to avoid ice formation in later steps. a spatula without introducing air bub
Slide 7: 2.2 Sample embedding steps
2.2 Sample embedding steps 5. Use mold B as a lid with opening facing up, place on top of mold A 6. After 5 min, remove mold B and check if the OCT is completely frozen and gently and then place a few dry ice cubes on top of mold B. Make sure turns opaque, otherwise repeat step 5. the two stainless-
Slide 8: 2.2 Sample embedding steps
2.2 Sample embedding steps 9. Label the tissue block to mark the orientation of the tissue: ensure that the tissue is in the correct orientation after embedding in OCT. Label the sectioning side with a ; cutting would be started from this side. 10. Wrap the OCT-embedded tissue block with aluminum fo
Slide 9: RIN measurement and H&E staining after OCT embedding
- It is recommended to measure RIN after OCT embedding*. - Total RNA can be extracted from 10-20 slices of 10 μm-thick tissue sections to determine the RIN value (RIN≥7 is required). - H&E staining of a tissue section of the same tissue block after OCT embedding is required to check sample condition
Slide 10: Tissue orientation and trimming to target region
- It is important to ensure that the tissue is in the correct orientation after embedding in OCT. - After the RIN determination and H&E staining steps in section 3, the client should trim the OCT-embedded tissue block until it is close to the desired region for analysis. The client should leave abou
Slide 11: Can multiple samples be mounted on one chip?
Can multiple samples be mounted on one chip? Yes, provided the multiple samples can be embedded on 1 cryomold and the samples are of the same tissue type, of similar size, and are embedded to the same plane. However, we recommend not to put more than 4 tissue samples on 1 cryomold to ensure adequate
Slide 12: Tissue QC: sectioning for RIN and H&E
Tissue QC: sectioning for both RIN and H&E Step 1: Trim off excess OCT and some tissue sections (min. 3-5 sections) to fully expose the tissue; cut 6x10µm sections for RIN check (customer will be notified on RIN value) Step 2: Cut 10µm for H&E - Tissue quality check by Mirxes; customer will be notif
Slide 13: If target region selection and/or ROI selection required
If target region selection and/or ROI selection required ROI selection if any Trim towards the target region (area/structure of interest within the tissue plane) Trim until about 200-300µm away from the target region, then cover with OCT We will take sections from here for RIN check and tissue Tissu
Slide 14: Section 7. Sample shipment
mirxes Section 7. Sample shipment - If there is plastic mold available, you can place the OCT block inside the mold (of a suitable size) for shipment to provide extra protection to the tissue block. Otherwise, you can choose to place the OCT block inside pre-cooled, sealed small containers to minimi
Mirxes Holding Company — Technical Presentation (April 2025) on 6ix