Rapid confocal imaging for measuring sound-induced motion of the hearing organ in the apical region.
We describe a novel confocal image acquisition system capable of measuring the sound-evoked motion of the organ of Corti. The hearing organ is imaged with a standard laser scanning confocal microscope during sound stimulation. The exact temporal relation between each image pixel and the sound stimulus is quantified. The motion of the structures under study is obtained by fitting a Fourier series to the time dimension of a continuous sequence of acquired images. Previous versions of this acquisition system used a simple search to find pixels with similar phase values. The Fourier series approach permits substantially faster image acquisition with reduced noise levels and improved motion estimation. The system is validated by imaging various vibrating samples attached to a feedback-controlled piezoelectric translator. When using a rigid sample attached to the translator, the system is capable of measuring motion with peak-to-peak amplitudes smaller than 50 nm with an error below 20% at frequencies between 50 and 600 Hz. Examples of image sequences from the inner ear are given, along with detailed performance characteristics of the method.
Jacob
Stefan
S
Karolinska Institutet, Center for Hearing and Communication Research, Departments of Otolaryngology and Clinical Neuroscience, M1 Karolinska Universitetssjukhuset, SE-171 76 Stockholm, Sweden.
Tomo
Igor
I
Fridberger
Anders
A
de Monvel
Jacques Boutet
JB
Ulfendahl
Mats
M
D000161:Acoustic Stimulation / Q000379:methods; D000818:Animals; D003199:Computer Systems; D005260:Female; D006168:Guinea Pigs; D006309:Hearing / Q000502:physiology*; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D040542:Mechanotransduction, Cellular / Q000502:physiology; D018613:Microscopy, Confocal / Q000379:methods*; D009068:Movement / Q000502:physiology*; D009925:Organ of Corti / Q000166:cytology* / Q000502:physiology*