Mechanical Cell Disruption Using Hierarchical Micro-Nano Structureson Nanoporous Alumina Filter
Notice bibliographique
Résumé
Cell disruption is essential process in molecular biology and biomedical engineering for investigation of DNA, RNA, virus and protein in cell. Generally, there are some methods for cell disruption or cell lysis such as chemical, electrical and mechanical method. [1, 2] Chemical cell lysis is method that dissolve cell envelope and nucleus using cell lysis buffer and lysate from this process contains lysis buffer which couldn't know the composition. Electrical and mechanical cell disruption methods required external equipment and shows low cell disruption efficiency. Especially, mechanical cell disruption is easy and quick method and studied using microstructures for increment of cell disruption efficiency. Hierarchical micro-nano structures on nanoporous alumina filter was fabricated aluminium wet etching process and multi-step anodic aluminium oxidation process. Pure aluminium sheet (5N, 1mm thickness) was mechanical and electro-polished for mirror-like surface. Next, aluminium sheet was dipped in aluminium etchant of CuCl 2 base for fabrication of micro-structures. During aluminium etching process, micro-structures were formed by chipping off a grain boundary of aluminium. Multi-step anodic aluminium oxidation process was carried out for fabrication of nano-structures and filter body. Aluminium that formed micro-structures on surface was anodized twice in 0.3M phosphoric acid aqueous solution of -5C under 180V. Formed nanoporous alumina structure was expanded by wet etching process for forming of spike-like nanostructure shape in 0.1M phosphoric acid aqueous solution of 35C. 3 rd anodic aluminium oxidation process was carried out for fabrication of filter body under same condition mentioned above for 40hours. Finally, aluminium was removed and barrier layer of nanoporous alumina was opened by wet etching process. For mechanical cell disruption, NIH3T3 fibroblast cells (1 10 6 cells/ml) in PBS (phosphate buffered saline) were injected through hierarchical micro-nano structures on nanoporous alumina filter which assembled with commercial filter holder by air pressure (5bar). Cell disruption efficiency was evaluated by quantification of DNA and protein in lysate. Concentration of DNA and protein were quantified using DNA isolation kit (DNeasy, Quiagen) and Bradford assay, respectively. According to results, 5.1 1.3 g/ml of DNA and 46 11 g/ml of protein were detected from lysate. In cell disruption process using hierarchical micro-nano structure on nanoporous alumina filter, cell envelope and nucleus were teared by micro-structures and un-teared nucleus by micro-structure was disrupted by nano-structures. This results shows cell envelop disruption of 32 % and cell nucleus disruption of 92 % compared to theoretical value. [7,
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Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,001 | 0,001 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».