Lung inflation with direct injection of agarose: a technique for simultaneous molecular and morphometric measurements.
The usual methods for preparing lungs for morphologic study involve the instillation of fixatives that modifyproteins and RNA such that the tissue is unsuitable for molecular studies. To develop a technique suitable for molecular studies, pieces of adult rat lungs were infiltrated with agarose, glutaraldehyde, or paraformaldehyde and the consistency of alveolar inflation was compared to lungs inflated with 10% formalin. Only direct injection with 1% agarose resulted in comparable inflation of lung tissue and preserved RNA and protein. Thus, this technique enables simultaneous molecular and morphometric analysis of the lung on small pieces of lung tissue in heterogeneous lung diseases.
Bowler
Russell P
RP
Department of Medicine, National Jewish Medical and Research Center, Denver, Colorado 80206, USA. BowlerR@njc.org
Ellison
Misoo C
MC
Duda
Beth
B
Tran
Karen
K
Nicks
Mike
M
Cool
Carlyne
C
Greene
Kelly
K
Crapo
James D
JD
D000818:Animals; D005557:Formaldehyde; D005976:Glutaral; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D007267:Injections; D008168:Lung; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D011108:Polymers; D011506:Proteins / Q000032:analysis; D011650:Pulmonary Alveoli / Q000033:anatomy & histology / Q000737:chemistry; D012313:RNA / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012685:Sepharose; D016707:Tissue Fixation / Q000379:methods*
D019188:Organ Preservation Solutions; D011108:Polymers; D011506:Proteins; D005557:Formaldehyde; D012313:RNA; D012685:Sepharose; D005976:Glutaral; C003043:paraform